Efflux pump inhibitors

ABSTRACT

Compounds are described which have efflux pump inhibitor activity. Also described are methods of using such efflux pump inhibitor compounds and pharmaceutical compositions which include such compounds.

FIELD OF THE INVENTION

This invention relates to the field of antimicrobial agents and tomethods for identification and characterization of potentialantimicrobial agents. More particularly, this invention relates toantimicrobial agents for which the mode of action involves cellularefflux pumps and the regulation of efflux pumps.

BACKGROUND

The following background material is not admitted to be prior art to thepending claims, but is provided only to aid the understanding of thereader.

Antibiotics have been effective tools in the treatment of infectiousdiseases during the last half century. From the development ofantibiotic therapy to the late 1980s there was almost complete controlover bacterial infections in developed countries. The emergence ofresistant bacteria, especially during the late 1980s and early 1990s, ischanging this situation. The increase in antibiotic resistant strainshas been particularly common in major hospitals and care centers. Theconsequences of the increase in resistant strains include highermorbidity and mortality, longer patient hospitalization, and an increasein treatment costs. (B. Murray, 1994, New Engl. J. Med. 330: 1229-1230.)

The constant use of antibiotics in the hospital environment has selectedbacterial populations that are resistant to many antibiotics. Thesepopulations include opportunistic pathogens that may not be stronglyvirulent but that are intrinsically resistant to a number ofantibiotics. Such bacteria often infect debilitated or immunocompromisedpatients. The emerging resistant populations also include strains ofbacterial species that are well known pathogens, which previously weresusceptible to antibiotics. The newly acquired resistance is generallydue to DNA mutations, or to resistance plasmids (R plasmids) orresistance-conferring transposons transferred from another organism.Infections by either type of bacterial population, naturally resistantopportunistic pathogens or antibiotic-resistant pathogenic bacteria, aredifficult to treat with current antibiotics. New antibiotic moleculeswhich can override the mechanisms of resistance are needed.

Bacteria have developed several different mechanisms to overcome theaction of antibiotics. These mechanisms of resistance can be specificfor a molecule or a family of antibiotics, or can be non-specific and beinvolved in resistance to unrelated antibiotics. Several mechanisms ofresistance can exist in a single bacterial strain, and those mechanismsmay act independently or they may act synergistically to overcome theaction of an antibiotic or a combination of antibiotics. Specificmechanisms include degradation of the drug, inactivation of the drug byenzymatic modification, and alteration of the drug target (B. G. Spratt,Science 264:388 (1994)). There are, however, more general mechanisms ofdrug resistance, in which access of the antibiotic to the target isprevented or reduced by decreasing the transport of the antibiotic intothe cell or by increasing the efflux of the drug from the cell to theoutside medium. Both mechanisms can lower the concentration of drug atthe target site and allow bacterial survival in the presence of one ormore antibiotics which would otherwise inhibit or kill the bacterialcells. Some bacteria utilize both mechanisms, combining a lowpermeability of the cell wall (including membranes) with an activeefflux of antibiotics. (H. Nikaido, Science 264:382-388 (1994)).

In some cases, antibiotic resistance due to low permeability is relatedto the structure of the bacterial membranes. In general, bacteria can bedivided into two major groups based on the structure of the membranessurrounding the cytoplasm. Gram-positive (G+) bacteria have onemembrane, a cytoplasmic membrane. In contrast, Gram-negative (G-)bacteria have two membranes, a cytoplasmic membrane and an outermembrane. These bacterial membranes are lipid bilayers which containproteins and may be associated with other molecules. The permeability ofbacterial membranes affects susceptibility/resistance to antibioticsbecause, while there are a few molecular targets of antibiotics, e.g.,penicillin-binding proteins, that are accessible from the outer leafletof the cytoplasmic membranes, the principal targets for antibiotics arein the cytoplasm or in the inner leaflet of the cytoplasmic membrane.Therefore for an antibiotic which has a target in the cytoplasmicmembrane, in Gram-negative bacteria that antibiotic will first need tocross the outer membrane. For a target in the cytoplasm, an antibioticwill need to cross the cytoplasmic membrane in Gram-positive bacteria,and both the outer and cytoplasmic membranes in Gram-negative bacteria.For both membranes, an antibiotic may diffuse through the membrane, ormay cross using a membrane transport system.

For Gram-negative bacteria, the lipid composition of the outer membraneconstitutes a significant permeability barrier. The outer layer of thisouter membrane contains a lipid, lipopolysaccharide (LPS), which is onlyfound in the outer membrane of Gram-negative bacteria. The lipid layerof the outer membrane is highly organized in a quasi-crystalline fashionand has a very low fluidity. Because of the low fluidity of the lipidlayer of the outer membrane, even lipophilic antibiotics will notdiffuse rapidly through the lipid layer. This has been shownexperimentally, hydrophobic probe molecules have been shown to partitionpoorly into the hydrophobic portion of LPS and to permeate across theouter membrane bilayer at about one-fiftieth to one-hundredth the ratethrough the usual phospholipid bilayers (like the cytoplasmic membranebilayer).

Some antibiotics may permeate through water-filled porin channels orthrough specific transport systems. Many of the porin channels, however,provide only narrow diameter channels which do not allow efficientdiffusion of the larger antibiotic molecules. In addition, many porinchannels are highly hydrophilic environments, and so do not efficientlyallow the passage of hydrophobic molecules. Thus, the outer membraneacts as a molecular sieve for small molecules. This explains, in part,why Gram-negative bacteria are generally less susceptible to antibioticsthan Gram-positive bacteria, and why Gram-negative bacteria aregenerally more resistant to large antibiotics, such as glycopeptides,that cannot cross the outer membrane.

The cytoplasmic membrane also provides a diffusion barrier for someantibiotics. However, since the fluidity of the lipid layer of thecytoplasmic membrane is higher than that of the outer membrane ofGram-negative bacteria, drugs that show some lipophilicity will be ableto permeate through the lipid layer. Other drugs, such as phosphonomycinor D-cycloserine that have very low solubility in a lipophilicenvironment will cross the cytoplasmic membrane by using a transportsystem. In this case, though, if the transport system is notsynthesized, the bacteria will become resistant to the drug (Peitz etal., 1967, Biochem. J. 6: 2561).

Decreasing the permeability of the outer membrane, by reducing eitherthe number of porins or by reducing the number of a certain porinspecies, can decrease the susceptibility of a strain to a wide range ofantibiotics due to the decreased rate of entry of the antibiotics intothe cells. However, for most antibiotics, the half-equilibration timesare sufficiently short that the antibiotic could exert its effect unlessanother mechanism is present. Efflux pumps are an example of such othermechanism. Once in the cytoplasm or periplasm a drug can be transportedback to the outer medium. This transport is mediated by efflux pumps,which are constituted of proteins. Different pumps can effluxspecifically a drug or group of drugs, such as the NorA system thattransports quinolones, or Tet A that transports tetracyclines, or theycan efflux a large variety of molecules, such as certain efflux pumps ofPseudomonas aeruginosa. In general, efflux pumps have a cytoplasmiccomponent and energy is required to transport molecules out of the cell.Some efflux pumps have a second cytoplasmic membrane protein thatextends into the periplasm. At least some efflux pumps of P. aeruginosahave a third protein located in the outer membrane.

Efflux pumps are involved in antibiotic resistance since, in some cases,they can remove a significant fraction of the antibiotic molecules whichmanage to enter the cells, thereby maintaining a very low intracellularantibiotic concentration. To illustrate, P. aeruginosalaboratory-derived mutant strain 799/61, which does not produce anymeasurable amounts of efflux pump is 8 to 10 fold more susceptible totetracycline and ciprofloxacin than the parent strain P. aeruginosa 799,which synthesizes efflux pumps. Also, null mutants of mexA, thecytoplasmic component of a P. aeruginosa efflux pump, are moresusceptible to antibiotics than the wild type.

The physiological role of efflux pumps has not been clearly defined yet.They are involved in drug resistance but they also are involved in thenormal physiology of the bacterial cell. The efflux pump coded in themexA operon of P. aeruginosa has been shown to be regulated by the ironcontent of the medium, and it is co-regulated with the synthesis of thereceptors of siderophores. Siderophores are molecules that are neededfor bacterial growth under iron starvation conditions, such as duringinfection of an animal. They are synthesized in the cytoplasm andexported when the bacterial cell needs iron. Siderophores scavenge ironwithin the infected animal and return the iron to the microbe to be usedfor essential microbial processes. Since there is essentially no freeiron in the bodies of animals, including the human body, the productionof siderophores by infecting bacteria is an important virulence factorfor the progress of the infection.

Even organisms normally surrounded by a cell envelope of relatively highpermeability can develop resistance by decreasing the permeability ofthe envelope. When an agent mainly diffuses across the barrier through aspecific channel, mutational loss of the channel can be an efficientmechanism for resistance. A "nonclassical" β-lactam compound, imipenem,shows an exceptional activity against P. aeruginosa, mainly because thisagent diffuses though a specific channel, OprD, whose physiologicalfunction appears to be that of the transport of basic amino acids.However, P. aeruginosa could become resistant to imipenem by simplylosing the oprD channel, and currently a large fraction of P. aeruginosastrains isolated from the hospital environment are resistant as a resultof this modification. In a similar manner, β-lactam compounds designedto mimic iron-chelating compounds (siderophores) during their transportthrough the outer membranes are known to select mutants that aredefective in the specific transport of these siderophores.

In summary, the above discussion indicates that cellular factorsaffecting transport (both active and passive transport) of antibioticsinto bacterial cells are important components of antibiotic resistancefor many bacterial species.

SUMMARY

This invention concerns particular compounds which are efflux pumpinhibitors, and which are therfore compounds which inhibit cellularefflux pumps of bacteria or other microbes. Such efflux pumps exportsubstrate molecules from the cytoplasm in an energy-dependent manner,and the exported substrate molecules can include antibacterial agents.Such efflux pump inhibitors are useful, for example, for treatingmicrobial infections by reducing the export of a co-administeredantimicrobial agent or by preventing the export of a compoundsynthesized by microbes (e.g., bacteria) to allow or improve theirgrowth. An example of reducing the export of such a compound isinhibiting iron availability for the microbe by reducing the export ofsiderophores. Thus, this invention also provides compositions whichinclude such efflux pump inhibitors and methods for treating microbialinfections using those compositions.

The identification and use of efflux pump inhibitors is described inco-pending United States patent applications, Trias et al., EFFLUX PUMPINHIBITORS, application Ser. No. 08/427,088, filed Apr. 21, 1995 andTrias et al., EFFLUX PUMP INHIBITORS, application Ser. No. 08/898,477,filed Jul. 22, 1997, which are hereby incorporated by reference in theirentireties including drawings. Screening methods described therein wereused to identity some of the efflux inhibitor compounds of the presentinvention, and additional compounds were synthesized and tested whichwere structurally related to the active compounds identified throughscreening.

The efflux pump inhibitors of the present invention have structureswhich are shown by the generic structure 1 below: ##STR1## whereM*═(CH₂)_(n) (n=0-2)

P*═CH₂, carbonyl (C═O), thiocarbonyl (C═S)

S*═CH₂, CH(OH), NH, O, SO_(t) (t=0-2);

R═H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═((NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b) or R^(c))═H, lower alkyl, phenyl,substituted phenyl, benzyl, cyano, hydroxyl, or nitro. AlternativelyR^(a) +R^(b) (or R^(b) +R^(c))═(CH₂)₂₋₃ or --CH═CH--.

R¹ ═H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b) or R^(c))═H, lower alkyl, phenyl,benzyl, cyano, hydroxyl, or nitro. Alternatively R^(a) +R^(b) (or R^(b)+R^(c))═(CH₂)₂₋₃ or --CH═CH--.

R² ═H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl, aryl,monosubstituted aryl, disubstituted aryl, 2-(or 3-)thienyl, 2-(or3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl, benzothienyl, indolyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, benzofuranylalkyl, benzothienylalkyl,indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), (CH₂)_(n) N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b) or R^(c))═H,alkyl, phenyl, benzyl, cyano, hydroxyl, or nitro. Alternatively R^(a)+R^(b) (or R^(b) +R^(c))═(CH₂)₂₋₃ or --CH═CH--.

W═(alpha-aminoacyl)amido (such as glycylamido, D-alanylamido,D-aspartylamido, D-glutamylamido, D-leucylamido, D-phenylalanylamido,D-phenylglycylamido, D-tyrosylamido), aminoalkyl [(CH₂)_(n) NR^(b) R^(c); n=1-4; R^(b) and/or R^(c) ═H, lower alkyl, aryl], amino,azaheterocycles [such as N-morpholinyl, N-piperazinyl, N-pyrrolidinyl,N-imidazolyl, N-pyrrolyl, N-pyrazolyl, N-triazolyl, or N-tetrazolyl],substituted azaheterocycles [e.g., 2-(or 3-) lower alkylmorpholinyl,2-(3- or 4-) lower alkylpiperazinyl, 2-(or 3-) lower alkylpyrrolidinyl,2-(or 3-) lower alkylmorpholinyl, 2-(or 3-) lower alkylpyrrolyl],hydroxyl, alkoxy, alkylthio, guanidino, amidino, or halogen.

X═aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,tetrahydronaphthyl, indanyl, quinolinyl, quinolinyl, isoquinolinyl,quinoxalinyl, quinazolinyl, benzimidazolyl, benzothiazolyl,benzoxazolyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,quinolinylalkyl, isoquinolinylalkyl, quinoxalinylalkyl,quinazolinylalkyl, benzimidazolylalkyl, benzothiazolylalkyl,benzoxazolylalkyl.

Where there are centers of asymmetry, the absolute stereochemistry canbe either R or S-configuration and any combination of configuration.Even racemic materials fulfill the structural generics descriptions.

In the generic descriptions of compounds of this invention, the numberof atoms of a particular type in a substituent group is generally givenas a range. For example, an alkyl group containing from 1 to 4 carbonatoms is indicated as alkyl (C₁ -C₄), or as (C₁₋₄) alkyl. Such a rangereference is intended to include specific references to groups havingeach of the integer number of atoms within the specified range. Forecample, C₁ -C₄ includes each of C₁, C₂, C₃ and C₄. Other numbers ofatoms and other types of atoms are indicated in a similar manner.

Unless otherwise indicated, the term "alkyl" refers to a branched orunbranched aliphatic hydrocarbon group, preferably having from 1 to 6carbon atoms, and more preferably 1 to 4 carbon atoms. Preferably thehydrocarbon group is saturated. The alkyl group may optionally besubstituted, and some preferred subsituents include alkoxy, alkylthio,halogen, amino, monosubstituted amino, disubstituted amino, and carboxygroups.

The term "lower alkyl" refers to an aliphatic hydrocarbon having 1 to 6carbons, and preferably 1 to 4 carbon atoms (i.e., 1, 2, 3, or 4 carbonatoms). The lower alkyl group may be substituted; preferred substituentsinclude alkoxy, alkylthio, halogen, amino, monosubstituted amino,disubstituted amino, and carboxy.

The term "branched alkyl" refers to a branched aliphatic hydrocarbon.The branched alkyl group is preferably 3 to 10 (i.e., 3, 4, 5, 6, 7, 8,9 or 10 carbon atoms) carbons, and most preferably 3 to 6 carbons (i.e.,3, 4, 5, or 6 carbon atoms). The branched alkyl group may be substitutedand some preferred substituents include alkoxy, alkylthio, halogen,amino, monosubstituted amino, disubstituted amino, and carboxy.

The term "fluoroalkyl" refers to a lower alkyl group which issubstituted with a fluorine. The term "perfluoroalkyl" refers to a loweralkyl group which is substituted with a fluorine atom in every availableposition except for where the lower alkyl group is attached to the mainchain.

The term "carboxyalkyl" refers to a chemical moiety with formula--(R)n--COOH, where R is an alkyl moiety, preferably a saturated alkyl,and where n is 0-5.

The term "hydroxyalkyl" refers to a chemical moiety with the formula--(R)n--OH, where R is an alkyl moiety and where n is 1-4.

The term "alkoxy" refers to a chemical substituent of formula --OR,where R is hydrogen or a saturated or unsaturated lower alkyl moiety.

The term "alkylthio" refers to a chemical substituent of formula --SR,where R is hydrogen or a saturated or unsaturated lower alkyl moiety.

The term "aryl" refers to an aromatic group which has at least one ringhaving a conjugated p electron system and includes both carbocyclic aryl(e.g. phenyl) and heterocyclic aryl groups (e.g. pyridine). The arylgroup is preferably 6 to 14 carbons, more preferably 6 to 10 carbons.Aryl moieties include monocyclic, bicyclic, and tricyclic rings, whereeach ring has preferably five or six members. The aryl moiety may beoptionally monosubstituted or disubstituted with lower alkyl, hydroxyl,alkoxy, alkylthio, halogen, amino, monosubstituted amino, anddisubstituted amino.

The term "carbocyclic" refers to a compound which contains one or morecovalently closed ring structures, and that the atoms forming thebackbone of the ring are all carbon atoms. The term thus distinguishescarbocyclic from heterocyclic rings in which the ring backbone containsat least one atom which is different from carbon.

Thus, the term "azaheterocycle" refers to a heterocyclic group whichincludes at least one nitrogen atom in a ring. Preferably theazaheterocyclic group is a N-morpholinyl, N-piperazinyl, N-pyrrolidinyl,N-imidazolyl, N-pyrrolyl, N-pyrazolyl, N-triazolyl, and N-tetrazolylgroup. The azaheterocyclic group may also be substituted as recognizedin the art, forming a substituted azaheterocycle, preferably a 2-(or 3-)lower alkylnorpholinyl, 2-(3- or 4-) lower alkylpiperazinyl, 2-(or 3-)lower alkylpyrrolidinyl, 2-(or 3-) lower alkylmorpholinyl, 2-(or 3-)lower alkylpyrrolyl group.

The term "monosubstituted aryl" refers to an aryl group substituted witha group selected from alkyl, alkoxy, alkylthio, halogen, hydroxyl,amino, monosubstituted amino, or disubstituted amino.

"Halogen" or "halo" refers to F, Br, Cl, or I, but is preferably F orBr, and more preferably is F.

"Hydroxyl" or "hydroxy" refers to the group --OH.

The term "amino" means the group NRR', where R and R' may independentlybe alkyl or hydrogen or hydroxyl, but preferably are hydrogen. The term"monosubstituted amino" refers to an amino group in which one of R or R'is alkyl. The term "disubstituted amino" refers to an amino group inwhich R and R' are each independently alkyl or hydroxyl.

The term "arylalkyl" refers to a lower alkyl group substituted with anaryl group. An example of an arylalkyl group is benzyl where a methylgroup is substituted with phenyl. The lower alkyl group may beoptionally substituted with a lower alkyl, alkoxy, alkylthio, halogen,amino, monosubstituted amino, or disubstituted amino. The arylalkylgroup may be aryl-substituted where the aryl group is optionallysubstituted with a lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino.

The term "thienylalkyl" refers to a lower alkyl group substituted with athienyl group. The lower alkyl group may be optionally substituted witha lower alkyl, alkoxy, alkylthio, halogen, amino, monosubstituted amino,or disubstituted amino. The thienylalkyl group may bethienyl-substituted where the thienyl group is optionally substitutedwith a lower alkyl, alkoxy, alkylthio, halogen, amino, monosubstitutedamino, or disubstituted amino.

The term "furylalkyl" refers to a lower alkyl group substituted with afuryl group. The lower alkyl group may be optionally substituted with alower alkyl, alkoxy, alkylthio, halogen, amino, monosubstituted amino,or disubstituted amino. The furylalkyl group may be furyl-substitutedwhere the furyl group is optionally substituted with a lower alkyl,alkoxy, alkylthio, halogen, amino, monosubstituted amino, ordisubstituted amino.

The term "pyridylalkyl" refers to a lower alkyl group substituted with apyridyl group. The lower alkyl group may be optionally substituted witha lower alkyl, alkoxy, alkylthio, halogen, amino, monosubstituted amino,or disubstituted amino. The pyridylalkyl group may bepyridyl-substituted where the pyridyl group is optionally substitutedwith a lower alkyl, alkoxy, alkylthio, halogen, amino, monosubstitutedamino, or disubstituted amino.

The term "benzothienylalkyl" refers to a lower alkyl group substitutedwith a benzothienyl group. The lower alkyl group may be optionallysubstituted with a lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino. The benzothienylalkylgroup may be benzothienyl-substituted where the benzothienyl group isoptionally substituted with a lower alkyl, alkoxy, alkylthio, halogen,amino, monosubstituted amino, or disubstituted amino.

The term "indolyalkyl" refers to a lower alkyl group substituted with anindole group. The lower alkyl group may be optionally substituted with alower alkyl, alkoxy, alkylthio, halogen, amino, monosubstituted amino,or disubstituted amino. The indolyalkyl group may be indole-substitutedwhere the indole group is optionally substituted with a lower alkyl,alkoxy, alkylthio, halogen, amino, monosubstituted amino, ordisubstituted amino.

The term "(alpha-aminoacyl)amido" refers to a group having an amidelinkage and which is alpha-amino substituted. Preferably the group is anamide-linked alpha-amino acid, which may optionally be substituted, forexample, glycylamido, D-alanylamido, D-aspartylamido, D-glutamylamido,D-leucylamido, D-phenylalanylamido, D-phenylglycylamido, D-tyrosylamido.

The term "aminoalkyl" refers to an amino substituted lower alkyl group,preferably (CH₂)_(n) NR^(b) R^(c) where n=1-4; R^(b) and/or R^(c) is H,lower alkyl, aryl.

The term "quinolinylalkyl" refers to a lower alkyl group substitutedwith an quinolinyl group. The lower alkyl group may be optionallysubstituted with a lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino. The quinolinylalkyl groupmay be quinolinyl-substituted where the quinolinyl group is optionallysubstituted with a lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino.

The term "isoquinolinylalkyl" refers to a lower alkyl group substitutedwith an isoquinolinyl group. The lower alkyl group may be optionallysubstituted with a lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino. The isoquinolinylalkylgroup may be isoquinolinyl-substituted where the quinolinyl group isoptionally substituted with a lower alkyl, alkoxy, alkylthio, halogen,amino, monosubstituted amino, or disubstituted amino.

The term "quinoxalinylalkyl" refers to a lower alkyl group substitutedwith an quinoxalinyl group. The lower alkyl group may be optionallysubstituted with a lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino. The quinoxalinylalkylgroup may be quinoxalinyl-substituted where the quinolinyl group isoptionally substituted with a lower alkyl, alkoxy, alkylthio, halogen,amino, monosubstituted amino, or disubstituted amino.

The term "quinazolinylalkyl" refers to a lower alkyl group substitutedwith an quinazolinyl group. The lower alkyl group may be optionallysubstituted with a lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino. The quinazolinylalkylgroup may be quinazolinyl-substituted where the quinazolinylgroup isoptionally substituted with a lower alkyl, alkoxy, alkylthio, halogen,amino, monosubstituted amino, or disubstituted amino.

The term "benzimidazolylalkyl" refers to a lower alkyl group substitutedwith an benzimidazolyl group. The lower alkyl group may be optionallysubstituted with a lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino. The benzimidazolylalkylgroup may be benzimidazolyl-substituted where the quinazolinylgroup isoptionally substituted with a lower alkyl, alkoxy, alkylthio, halogen,amino, monosubstituted amino, or disubstituted amino.

The term "benzothiazolylalkyl" refers to a lower alkyl group substitutedwith an benzothiazolyl group. The lower alkyl group may be optionallysubstituted with a lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino. The benzothiazolylalkylgroup may be benzothiazolyl-substituted where the quinazolinylgroup isoptionally substituted with a lower alkyl, alkoxy, alkylthio, halogen,amino, monosubstituted amino, or disubstituted amino.

The term "benzoxazolylalkyl" refers to a lower alkyl group substitutedwith an benzoxazolyl group. The lower alkyl group may be optionallysubstituted with a lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino. The benzoxazolylalkylgroup may be benzoxazolyl-substituted where the benzoxazolyl group isoptionally substituted with a lower alkyl, alkoxy, alkylthio, halogen,amino, monosubstituted amino, or disubstituted amino.

The term "benzofuranyl" refers to a group which has the core ringstructure of Structure A. The benzofuranyl group may be optionallysubstituted with lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino.

The term "benzothienyl" refers to a group which has the core ringstructure of Structure B. The benzothienyl group may be optionallysubstituted with lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino.

The term "indolyl" refers to a group which has the core ring structureof Structure C. The indolyl group may be optionally substituted withlower alkyl, alkoxy, alkylthio, halogen, amino, monosubstituted amino,or disubstituted amino.

The term "benzimidazolyl" refers to a group which has the core ringstructure of Structure D. The benzimidazolyl group may be optionallysubstituted with lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino.

The term "benzothiazolyl" refers to a group which has the core ringstructure of Structure E. The benzothiazolyl group may be optionallysubstituted with lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino.

The term "benzoxazolyl" refers to a group which has the core ringstructure of Structure F. The benzoxazolyl group may be optionallysubstituted with lower alkyl, alkoxy, alkylthio, halogen, amino,monosubstituted amino, or disubstituted amino. ##STR2##

In preferred embodiments, certain efflux pump inhibitors of the presentinvention have structures which are shown by the generic structure 2below: ##STR3## wherein R═H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, carboxyalkyl, hydroxyalkyl, aryl, monosubstituted aryl,disubstituted aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n)NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b) or R^(c))═H, lower alkyl, phenyl,benzyl, cyano, hydroxy, or nitro. Alternatively R^(a) +R^(b) (or R^(b)+R^(c))═(CH₂)₂₋₃ or --CH═CH--;

R¹ ═H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b) or R^(c))═H, lower alkyl, phenyl,benzyl, cyano, hydroxy, or nitro. Alternatively R^(a) +R^(b) (or R^(b)+R^(c))═(CH₂)₂₋₃ or --CH═CH--;

R² ═H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl, aryl,2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl,benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, benzofuranylalkyl,benzothienylalkyl, indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b)or R^(c))═H, lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro.Alternatively R^(a) +R^(b) (or R^(b) +R^(c))═(CH₂)₂₋₃ or --CH═CH--;

W═(alpha-aminoacyl)amido (such as glycylamido, D-alanylamido,D-aspartylamido, D-glutamylamido, D-leucylamido, D-phenylalanylamido,D-phenylglycylamido, or D-tyrosyl-amido), aminoalkyl [(CH₂)_(n) NR^(b)R^(c) ; n=1-4; R^(b) and/or R^(c) ═H, lower alkyl, aryl], amino,azaheterocycles (such as N-morpholinyl, N-piperazinyl, N-pyrrolidinyl,N-imidazolyl, N-pyrrolyl, N-pyrazolyl, N-triazolyl, or N-tetrazolyl),substituted azaheterocycles (such as 2-(or 3-) lower alkylmorpholinyl,2-(3- or 4-) lower alkylpiperazinyl, 2-(or 3-) lower alkylpyrrolidinyl,2-(or 3-) lower alkylmorpholinyl, 2-(or 3-) lower alkylpyrrolyl),hydroxy, alkoxy, alkylthio, guanidino, amidino, or halogen;

X═aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,tetrahydronaphthyl, indanyl, quinolinyl, isoquinolinyl, quinoxalinyl,quinazolinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, quinolinylalkyl,isoquinolinylalkyl, isoquinolinyl, quinoxalinylalkyl, quinazolinylalkyl,benzimidazolylalkyl, benzothiazolylalkyl, benzoxazolylalkyl;

where there are centers of asymmetry, the absolute stereochemistry canbe either R or S-configuration and any combination of configuration;even racemic materials fulfill the structural generics description.

In preferred embodiments, certain efflux pump inhibitors of the presentinvention have structures which are shown by the generic structure 3below: ##STR4## wherein R═H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, carboxy-alkyl, hydroxyalkyl, aryl, monosubstituted aryl,disubstituted aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n)NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b) or R^(c))═H, lower alkyl, phenyl,benzyl, cyano, hydroxy, or nitro. Alternatively R^(a) +R^(b) (or R^(b)+R^(c))═(CH₂)₂₋₃ or --CH═CH--

R¹ ═H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxy-alkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b) or R^(c))═H, lower alkyl, phenyl,benzyl, cyano, hydroxy, or nitro. Alternatively R^(a) +R^(b) (or R^(b)+R^(c))═(CH₂)₂₋₃ or --CH═CH--;

R² ═H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl, aryl,2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl,benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, benzofuranylalkyl,benzothienylalkyl, indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b)or R^(c))═H, lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro.Alternatively R^(a) +R^(b) (or R^(b) +R^(c))═(CH₂)₂₋₃ or --CH═CH--;

W═(alpha-aminoacyl)amido (such as glycylamido, D-alanylamido,D-aspartylamido, D-glutamylamido, D-leucylamido, D-phenylalanylamido,D-phenylglycylamido, or D-tyrosyl-amido), aminoalkyl ((CH₂)_(n) NR^(b)R^(c) ; n=1-4; R^(b) and/or R^(c) ═H, lower alkyl, aryl), amino,azaheterocycles (such as N-morpholinyl, N-piperazinyl, N-pyrrolidinyl,N-imidazolyl, N-pyrrolyl, N-pyrazolyl, N-triazolyl, or N-tetrazolyl),substituted azaheterocycles (such as 2-(or 3-) alkylmorpholinyl, 2-(3-or 4-) lower alkylpiperazinyl, 2-(or 3-) lower alkylpyrrolidinyl, 2-(or3-) lower alkylmorpholinyl, 2-(or 3-) lower alkylpyrrolyl), hydroxy,alkoxy, alkylthio, guanidino, amidino, or halogen;

X═aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,tetrahydronaphthyl, indanyl, quinolinyl, isoquinolinyl, quinoxalinyl,quinazolinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, quinolinylalkyl,isoquinolinylalkyl, isoquinolinyl, quinoxalinylalkyl, quinazolinylalkyl,benzimidazolylalkyl, benzothiazolylalkyl, benzoxazolylalkyl;

where there are centers of asymmetry, the absolute stereochemistry canbe either R or S-configuration and any combination of configuration;even racemic materials fulfill the structural generics description.

In preferred embodiments, certain efflux pump inhibitors of the presentinvention also have structures which are shown by the generic structure4 below: ##STR5## wherein S*═CH₂, CH(OH), NH, O, SO, (t=0-2);

R═H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, hydroxyalkyl, aryl, monosubstituted aryl, disubstitutedaryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b)R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c)(n=1-4); R^(a) (R^(b) or R^(c))═H, alkyl, phenyl, benzyl, cyano,hydroxy, or nitro. Alternatively R^(a) +R^(b) (or R^(b) +R^(c))═(CH₂)₂₋₃or --CH═CH--

R¹ ═H, alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl,, aryl,2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n)NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b) or R^(c))═H, lower alkyl, phenyl,benzyl, cyano, hydroxy, or nitro. Alternatively R^(a) +R^(b) (or R^(b)+R^(c))═(CH₂)₂₋₃ or --CH═CH--;

R² ═H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl, aryl,2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl,benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, benzofuranylalkyl,benzothienylalkyl, indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c) (n=1-4); R^(a) (R^(b)or R^(c))═H, lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro.Alternatively R^(a) +R^(b) (or R^(b) +R^(c))═(CH₂)₂₋₃ or --CH═CH--;

W═(alpha-aminoacyl)amido (such as glycylamido, D-alanylamido,D-aspartylamido, D-glutamylamido, D-leucylamido, D-phenylalanylamido,D-phenylglycylamido, or D-tyrosyl-amido), aminoalkyl ((CH₂)_(n) NR^(b)R^(c) ; n=1-4; R^(b) and/or R^(c) ═H, lower alkyl, aryl), amino,azaheterocycles (such as N-morpholinyl, N-piperazinyl, N-pyrrolidinyl,N-imidazolyl, N-pyrrolyl, N-pyrazolyl, N-triazolyl, or N-tetrazolyl),substituted azaheterocycles (such as 2-(or 3-) alkylmorpholinyl, 2-(3-or 4-) lower alkylpiperazinyl, 2-(or 3-) lower alkylpyrrolidinyl, 2-(or3-) lower alkylmorpholinyl, 2-(or 3-) lower alkylpyrrolyl], hydroxy,alkoxy, alkylthio, guanidino, amidino, or halogen;

X═aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,tetrahydronaphthyl, indanyl, quinolinyl, isoquinolinyl, quinoxalinyl,quinazolinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, quinolinylalkyl,isoquinolinylalkyl, quinoxalinylalkyl, quinazolinylalkyl,benzimidazolylalkyl, benzothiazolylalkyl, benzoxazolylalkyl;

where there are centers of asymmetry, the absolute stereochemistry canbe either R or S-configuration and any combination of configuration;even racemic materials fulfill the structural generics description.

In preferred embodiments of structure 2 compounds, the group R² isdifferent from hydrogen.

The generic compound descriptions above should be understood to includeadditional narrower generic descriptions in which the possiblesubstituents for one or more of the specified substituent groups orsubsitutions (e.g., W, R, R¹, R², X, M*, P*, S*) is limited to a subsetof the listed groups.

Compounds within the generic description above can be obtained bysynthetic chemistry methods known to those skilled in the chemical artsas exemplified in the Examples below. Specific compound examples withinthe generic description are provided in the Detailed Description belowin connection with Tables 1-4.

A particularly appropriate example of a microbe appropriate for the useof an efflux pump inhibitor is a pathogenic bacterial species,Pseudomonas aeruginosa, which is intrinsically resistant to many of thecommonly used antibacterial agents. Exposing this bacterium to an effluxpump inhibitor can significantly slow the export of an antibacterialagent from the interior of the cell or the export of siderophores.Therefore, if another antibacterial agent is administered in conjunctionwith the efflux pump inhibitor, the antibacterial agent, which wouldotherwise be maintained at a very low intracellular concentration by theexport process, can accumulate to a concentration which will inhibit thegrowth of the bacterial cells. This growth inhibition can be due toeither bacteriostatic or bactericidal activity, depending on thespecific antibacterial agent used. While P. aeruginosa is an example ofan appropriate bacterium, other bacterial and microbial species maycontain similar broad substrate pumps, which actively export a varietyof antimicrobial agents, and thus can also be appropriate targets.

In addition as suggested above, for some microbial, e.g., bacterial,species, efflux pump inhibitors can decrease the virulence of themicrobe, for example, by inhibiting the transport of factors importantfor pathogenicity. Again using P. aeruginosa as an example, inhibitionof an efflux pump in this bacterium inhibits the uptake of iron, whichis important for pathogenicity. The mechanism of bacterial irontransport involves molecules called siderophores, which are synthesizedand exported by bacterial cells via efflux pumps. These siderophoresbind tightly to iron scavenged from the host, and are then taken up bythe bacteria. In this way, the iron needed for bacterial metabolism isobtained, and an infection can be maintained.

Therefore, illustrating the utility of efflux pump inhibitors,inhibiting the efflux pump of P. aeruginosa allows obtaining one or moreof the following biological effects:

1. P. aeruginosa strains will become susceptible to antibiotics thatcould not be used for treatment of pseudomonad infections, or becomemore susceptible to antibiotics which do inhibit pseudomonal growth.

2. P. aeruginosa strains will become more susceptible to antibioticscurrently used for treatment of pseudomonad infections.

3. Virulence of P. aeruginosa will be attenuated because theavailability of iron will be hampered.

4. The inhibition of the pump or of one of the components of the pumpmay be lethal or prevent growth.

Obtaining even one of these effects provides a potential therapeutictreatment for infections by this bacterium. Also, as previouslymentioned, similar pumps are found in other microorganisms. Some or allof the above effects can also be obtained with those microbes, and theyare therefore also appropriate targets for detecting or using effluxpump inhibitors. Thus, the term "microbes" include, for example,bacteria, fungi, yeasts, and protozoa.

As indicated, the bacterium to be inhibited through the use of an effluxpump inhibitor can be from other bacterial groups or species, such asone of the following:

Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonasacidovorans, Pseudomonas alcaligenes, Pseudomonas putida,Stenotrophomonas maltophilia, Burkholderia cepacia, Aeromonashydrophilia, Escherichia coli, Citrobacter freundii, Salmonellatyphimurium, Salmonella typhi, Salmonella paratyphi, Salmonellaenteritidis, Shigella dysenteriae, Shigella flexneri, Shigella sonnei,Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae,Klebsiella oxytoca, Serratia marcescens, Francisella tularensis,Morganella morganii, Proteus mirabilis, Proteus vulgaris, Providenciaalcalifaciens, Providencia rettgeri, Providencia stuartii, Acinetobactercalcoaceticus, Acinetobacter haemolyticus, Yersinia enterocolitica,Yersinia pestis, Yersinia pseudotuberculosis, Yersinia intermedia,Bordetella pertussis, Bordetella parapertussis, Bordetellabronchiseptica, Haemophilus influenzae, Haemophilus parainfluenzae,Haemophilus haemolyticus, Haemophilus parahaemolyticus, Haemophilusducreyi, Pasteurella multocida, Pasteurella haemolytica, Branhamellacatarrhalis, Helicobacter pylori, Campylobacter fetus, Campylobacterjejuni, Campylobacter coli, Borrelia burgdorferi, Vibrio cholerae,Vibrio parahaemolyticus, Legionella pneumophila, Listeria monocytogenes,Neisseria gonorrhoeae, Neisseria meningitidis, Kingella, Moraxella,Gardnerella vaginalis, Bacteroides fragilis, Bacteroides distasonis,Bacteroides 3452A homology group, Bacteroides vulgatus, Bacteroidesovalus, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroideseggerthii, Bacteroides splanchnicus, Clostridium difficile,Mycobacterium tuberculosis, Mycobacterium avium, Mycobacteriumintracellulare, Mycobacterium leprae, Corynebacterium diphtheriae,Corynebacterium ulcerans, Streptococcus pneumoniae, Streptococcusagalactiae, Streptococcus pyogenes, Enterococcus faecalis, Enterococcusfaecium, Staphylococcus aureus, Staphylococcus epidermidis,Staphylococcus saprophyticus, Staphylococcus intermedius, Staphylococcushyicus subsp. hyicus, Staphylococcus haemolyticus, Staphylococcushominis, Staphylococcus saccharolyticus.

The term "efflux pump" refers to a protein assembly which exportssubstrate molecules from the cytoplasm or periplasm of a cell, in anenergy dependent fashion. Thus an efflux pump will typically be locatedin the cytoplasmic membrane of the cell (spanning the cytoplasmicmembrane). In Gram-negative bacteria the pump may span the periplasmicspace and there may also be portion of the efflux pump which spans theouter membrane. Certain efflux pumps will include a polypeptide whichhas at least 50% amino acid sequence similarity with a polypeptide whichis part of the Pseudomonas aeruginosa mexA/mexB/oprM efflux pump or theefflux pump overexpressed by P. aeruginosa Strain K385, or the effluxpump overexpressed by P. aeruginosa Strain PA04098E. Due to thedescribed sequence similarity of a component polypeptide of the effluxpump, such an efflux pump is termed a Pseudomonas aeruginosa-type effluxpump.

The term "non-tetracycline-specific efflux pump" refers to an effluxpump which is not highly specific for tetracycline (relative to otherantibiotics) and thus is not a tetracycline (tetracycline-specific)efflux pump. The term thus includes broad substrate pumps (efflux anumber of compounds with varying structural characteristics) and pumpswhich are highly specific for compounds (including antibiotics) otherthan tetracyclines. Tetracycline efflux pumps are involved in specificresistance to tetracycline in bacteria. (Speer et al., 1992, Clin.Microbiol. Rev. 5: 387-399.) As noted, these pumps are highly specificfor tetracyclines, and their presence confers high tetracyclineresistance to the cell. However, they do not confer resistance to otherantibiotics. The genes for the tetracycline pump components are found inplasmids in Gram-negative as well as in Gram-positive bacteria and canbe divided in two main groups, tetA(A-E), and tetK and tetL. TetA-Etetracycline resistance determinants contain a structural gene, tetA,which is a tetracycline specific pump, and a repressor gene, tetR, thatmediates inducible resistance to tetracyclines. Tetracycline effluxpumps belonging to this group are designated tetA(A), tetA(B), tetA(D),and tetA(E), and are found in Enterobacteriaceae and other Gram-negativebacteria. TetK and TetL are pumps involved in tetracycline resistance inGram-positive bacteria. The genes are regulated via translationalattenuation and are not homologous to tetA group.

An "efflux pump inhibitor" is a compound which specifically interfereswith the ability of an efflux pump to export its normal substrate, orother compounds such as an antibiotic. The inhibitor may have intrinsicantimicrobial (e.g., antibacterial) activity of its own, but at least asignificant portion of the relevant activity is due to the efflux pumpinhibiting activity. Of particular interest in this invention, arecompounds which inhibit the export or activity of efflux pumps whichhave a broad substrate range which includes antibacterial agents. Theterm "non-tetracycline-specific efflux pump inhibitor" refers to anefflux pump inhibitor which inhibits a non-tetracycline-specific effluxpump. The term "Pseudomonas aeruginosa-type efflux pump inhibitor"refers to an efflux pump inhibitor which inhibits a Pseudomonasaeruginosa-type efflux pump. A "Pseudomonas aeruginosa efflux pumpinhibitor" is an efflux pump inhibitor which inhibits the exportactivity of an efflux pump found in Pseudomonas aeruginosa.

By "comprising" it is meant including, but not limited to, whateverfollows the word "comprising". Thus, use of the term "comprising"indicates that the listed elements are required or mandatory, but thatother elements are optional and may or may not be present. By"consisting of" is meant including, and limited to, whatever follows thephrase "consisting of". Thus, the phrase "consisting of" indicates thatthe listed elements are required or mandatory, and that no otherelements may be present. By "consisting essentially of" is meantincluding any elements listed after the phrase, and limited to otherelements that do not interfere with or contribute to the activity oraction specified in the disclosure for the listed elements. Thus, thephrase "consisting essentially of" indicates that the listed elementsare required or mandatory, but that other elements are optional and mayor may not be present depending upon whether or not they affect theactivity or action of the listed elements.

In another aspect, this invention provides a method for treating amicrobial infection, e.g., a bacterial infection, in an animal byadministering to an animal suffering from such an infection an effluxpump inhibitor as described above in an amount sufficient to reduceefflux pump activity.

In a preferred embodiment, the inhibitor is one which decreases thepathogenicity of the microbe. Such a decrease in pathogenicity can beobtained, for example, by interfering with bacterial iron acquisition byinhibiting the transport of siderophores. The pathogenicity may also bereduced by reducing or eliminating the microbial products which causetissue-damaging effects to the host. Other methods of reducingpathogenicity are, however, also within this aspect. The animal may be,for example, chickens and turkeys, and in certain preferred embodimentsis a mammal, e.g., a human.

In certain preferred embodiments, the microbial infection may be due tobacteria, which may, for example, be any of the bacterial speciesindicated above, but specifically including Pseudomonas aeruginosa.

In a related aspect, this invention provides a method of treating ananimal suffering from a microbial infection by administering to theanimal an efflux pump inhibitor in an amount sufficient to reduce effluxpump activity. In this aspect, the efflux pump inhibitor in one whichreduces the in vivo viability of a microbe involved in the infection. Byreducing the in vivo viability, the infected animal can more readilyclear its body of the infection, or the microbes may even be killed. Inparticular embodiments the animal is a mammal. Also in particularembodiments, the microbe may be from one of a variety of pathogenicbacterial species, specifically including those listed above.

The term "in vivo viability" refers to the ability of a microbe, e.g., abacterium, to survive or grow in a host, such as an animal. Therefore,an efflux pump inhibitor which reduces the in vivo viability of amicrobe may stop the growth of the microbe and/or kill the microbe. Suchefflux pump inhibitors, therefore are antimicrobial agents.

In a further related aspect, this invention includes a method forprophylactic treatment of an animal, e.g., a mammal. In this method, anefflux pump inhibitor which reduces the pathogenicity of a microbe isadministered to a mammal at risk of a microbial infection, e.g., abacterial infection.

In a related aspect, the invention provides a method for treating amicrobial infection in an animal, specifically including in a mammal, bytreating an animal suffering from such an infection with anantimicrobial agent and an efflux pump inhibitor which increase thesusceptibility of the microbe for that antimicrobial agent. In this waya microbe involved in the infection can be treated using theantimicrobial agent in smaller quantities, or can be treated with anantimicrobial agent which is not therapeutically effective when used inthe absence of the efflux pump inhibitor. Thus, this method of treatmentis especially appropriate for the treatment of infections involvingmicrobial strains which are difficult to treat using an antimicrobialagent alone due to a need for high dosage levels (which can causeundesirable side effects), or due to lack of any clinically effectiveantimicrobial agents. However, it is also appropriate for treatinginfections involving microbes which are susceptible to particularantimicrobial agents as a way to reduce the dosage of those particularagents. This can reduce the risk of side effects, but can also reducethe selection effect for highly resistant microbes resulting from theconsistent high level use of a particular antimicrobial agent. Inparticular embodiments the microbe is a bacterium, which may, forexample, be from any of the groups or species indicated above. Also inparticular embodiments various antibacterial agents can be used. Theseinclude quinolones, tetracyclines, glycopeptides, aminoglycosides,β-lactams, rifamycins, coumermycins, macrolides, and chloramphenicol. Inparticular embodiments an antibiotic of the above classes can be, forexample, one of the following:

β-Lactam Antibiotics

imipenem, meropenem, biapenem, cefaclor, cefadroxil, cefamandole,cefatrizine, cefazedone, cefazolin, cefixime, cefinenoxime, cefodizime,cefonicid, cefoperazone, ceforanide, cefotaxime, cefotiam, cefpimizole,cefpiramide, cefpodoxime, cefsulodin, ceftazidime, cefteram, ceftezole,ceftibuten, ceftizoxime, ceftriaxone, cefuroxime, cefuzonam,cephaacetrile, cephalexin, cephaloglycin, cephaloridine, cephalothin,cephapirin, cephradine, cefinetazole, cefoxitin, cefotetan, azthreonam,carumonam, flomoxef, moxalactam, amidinocillin, amoxicillin, ampicillin,azlocillin, carbenicillin, benzylpenicillin, carfecillin, cloxacillin,dicloxacillin, methicillin, mezlocillin, nafcillin, oxacillin,penicillin G, piperacillin, sulbenicillin, temocillin, ticarcillin,cefditoren, SC004, KY-020, cefdinir, ceftibuten, FK-312, S-1090,CP-0467, BK-218, FK-037, DQ-2556, FK-518, cefozopran, ME1228, KP-736,CP-6232, Ro 09-1227, OPC-20000, LY206763

Macrolides

azithromycin, clarithromycin, erythromycin, oleandomycin, rokitamycin,rosaramicin, roxithromycin, troleandomycin

Quinolones

amifloxacin, cinoxacin, ciprofloxacin, enoxacin, fleroxacin, flumequine,lomefloxacin, nalidixic acid, norfloxacin, ofloxacin, levofloxacin,oxolinic acid, pefloxacin, rosoxacin, temafloxacin, tosufloxacin,sparfloxacin, clinafloxacin, PD131628, PD138312, PD140248, Q-35, AM-1155, NM394, T-3761, rufloxacin, OPC-17116, DU-6859a (identified in Sato,K. et al., 1992, Antimicrob Agents Chemother. 37:1491-98), DV-7751a(identified in Tanaka, M. et al., 1992, Antimicrob. Agents Chemother.37:2212-18)

Tetracyclines

chlortetracycline, demeclocycline, doxycycline, lymecycline,methacycline, minocycline, oxytetracycline, tetracycline

Aminoglycosides

amikacin, arbekacin, butirosin, dibekacin, fortimicins, gentamicin,kanamycin, meomycin, netilmicin, ribostamycin, sisomicin, spectinomycin,streptomycin, tobramycin, clindamycin, lincomycin

In a further related aspect, this invention includes a method forprophylactic treatment of a mammal. In this method, an antimicrobialagent and an efflux pump inhibitor is administered to a mammal at riskof a microbial infection, e.g., a bacterial infection.

In the context of the response of a microbe, such as a bacterium, to anantimicrobial agent, the term "susceptibility" refers to the sensitivityof the microbe for the presence of the antimicrobial agent. So, toincrease the susceptibility means that the microbe will be inhibited bya lower concentration of the antimicrobial agent in the mediumsurrounding the microbial cells. This is equivalent to saying that themicrobe is more sensitive to the antimicrobial agent. In most cases theminimum inhibitory concentration (MIC) of that antimicrobial agent willhave been reduced.

As used herein, the term "treating" refers to administering apharmaceutical composition for prophylactic and/or therapeutic purposes.The term "prophylactic treatment" refers to treating a patient who isnot yet infected, but who is susceptible to, or otherwise at risk of, aparticular infection. The term "therapeutic treatment" refers toadministering treatment to a patient already suffering from aninfection. Thus, in preferred embodiments, treating is theadministration to a mammal (either for therapeutic or prophylacticpurposes) of therapeutically effective amounts of a potentiator and anantibacterial (or antimicrobial) agent in combination (eithersimultaneously or serially).

By "therapeutically effective amount" or "pharmaceutically effectiveamount" is meant an amount of an efflux pump inhibitor, or amountsindividually of an efflux pump inhibitor and an antimicrobial agent, asdisclosed for this invention, which have a therapeutic effect, whichgenerally refers to the inhibition to some extent of the normalmetabolism of microbial cells causing or contributing to a microbialinfection. The doses of efflux pump inhibitor and antimicrobial agentwhich are useful in combination as a treatment are therapeuticallyeffective amounts. Thus, as used herein, a therapeutically effectiveamount means those amounts of efflux pump inhibitor and antimicrobialagent which, when used in combination, produce the desired therapeuticeffect as judged by clinical trial results and/or model animal infectionstudies. In particular embodiments, the efflux pump inhibitor andantimicrobial agent are combined in pre-determined proportions and thusa therapeutically effective amount would be an amount of thecombination. This amount and the amount of the efflux pump inhibitor andantimicrobial agent individually can be routinely determined by one ofskill in the art, and will vary, depending on several factors, such asthe particular microbial strain involved and the particular efflux pumpinhibitor and antimicrobial agent used. This amount can further dependupon the patient's height, weight, sex, age and medical history. Forprophylactic treatments, a therapeutically effective amount is thatamount which would be effective if a microbial infection existed.

A therapeutic effect relieves, to some extent, one or more of thesymptoms of the infection, and includes curing an infection. "Curing"means that the symptoms of active infection are eliminated, includingthe elimination of excessive members of viable microbe of those involvedin the infection. However, certain long-term or permanent effects of theinfection may exist even after a cure is obtained (such as extensivetissue damage).

The term "microbial infection" refers to the invasion of the host mammalby pathogenic microbes. This includes the excessive growth of microbeswhich are normally present in or on the body of a mammal. Moregenerally, a microbial infection can be any situation in which thepresence of a microbial population(s) is damaging to a host mammal.Thus, a mammal is "suffering" from a microbial infection when excessivenumbers of a microbial population are present in or on a mammal's body,or when the effects of the presence of a microbial population(s) isdamaging the cells or other tissue of a mammal. Specifically, thisdescription applies to a bacterial infection.

The term "administration" or "administering" refers to a method ofgiving a dosage of an antimicrobial pharmaceutical composition to amammal, where the method is, e.g., topical, oral, intravenous,intraperitoneal, or intramuscular. The preferred method ofadministration can vary depending on various factors, e.g., thecomponents of the pharmaceutical composition, the site of the potentialor actual bacterial infection, the microbe involved, and the severity ofan actual microbial infection.

The term "mammal" is used in its usual biological sense. Thus, itspecifically includes humans, cattle, horses, dogs, and cats, but alsoincludes many other species.

In another aspect, this invention also features a method of inhibiting amembrane channel in a cellular membrane, involving contacting themembrane channel with a membrane channel inhibitor, where the inhibitorreduces the effluxing capacity of the membrane channel. In specificembodiments, at least one polypeptide of the membrane channel has atleast 50% amino acid sequence similarity with a polypeptide of themexA/mexB/oprM efflux pump, or of the efflux pump overexpressed byPseudomonas aeruginosa Strain K385.

As used herein, the term "membrane channel" refers to a protein assemblylocated in the cellular membrane of a cell which allows the transport ofone or more types of molecules across the membrane. Such transport maybe either passive transport in response to concentration gradients, ormay be active transport which depends upon a cellular energy source.

A "membrane channel inhibitor" then is, similar to an efflux pumpinhibitor, a compound which slows or prevents the transport of moleculesacross the cellular membrane using the corresponding membrane channel.

This invention also features a method of enhancing the antimicrobialactivity of an antimicrobial agent against a microbe, in which such amicrobe is contacted with an efflux pump inhibitor, e.g., anon-tetracycline specific efflux pump inhibitor, to an efflux pump inthe cell, and an antibacterial agent. The efflux pump inhibitor is acompound as described above. Thus, this method makes an antimicrobialagent more effective against a cell which expresses an efflux pump whenthe cell is treated with the combination of an antimicrobial agent and anon-tetracycline-specific efflux pump inhibitor. In particularembodiments the microbe is a bacterium or a fungus, such as any of thoseindicated in the first aspect above; the antibacterial agent can beselected from a number of structural classes of antibiotics including,e.g., β-lactams, glycopeptides, aminoglycosides, quinolones,tetracyclines, rifamycins, coumermycins, macrolides, andchloramphenicol. In particular embodiments an antibiotic of the aboveclasses can be as stated above.

In a further aspect this invention provides pharmaceutical compositionseffective for treatment of an infection of an animal, e.g., a mammal, bya microbe, such as a bacterium or a fungus. The composition includes apharmaceutically acceptable carrier and an efflux pump inhibitor asdescribed above. In preferred embodiments, such compositions containefflux pump inhibitors which are themselves effective antimicrobialagents, even in the absence of another antimicrobial agent (i.e., haveintrinsic antimicrobial activity). Thus, pharmaceutical compositionsincluding such efflux pump inhibitors can be used either alone or inconjunction with another antimicrobial agent. Also in preferredembodiments, the efflux pump inhibitors in pharmaceutical compositionsof this aspect are efflux pump inhibitors which enhance theeffectiveness of an animicrobial agent other than the efflux pumpinhibitor, so such compositions would generally be used in combinationwith such other antimicrobial agent. The invention also providespharmaceutical compositions similarly effective for treatment of aninfection of a mammal which include an efflux pump inhibitor and anantimicrobial agent. Similarly, the invention provides antimicrobialformulations which include an antimicrobial agent, an efflux pumpinhibitor, and a carrier. In preferred embodiments, the antimicrobialagent is an antibacterial agent.

A "carrier" or "excipient" is a compound or material used to facilitateadministration of the compound, for example, to increase the solubilityof the compound. Solid carriers include, e.g., starch, lactose,dicalcium phosphate, sucrose, and kaolin. Liquid carriers include, e.g.,sterile water, saline, buffers, non-ionic surfactants, and edible oilssuch as oil, peanut and sesame oils. In addition, various adjuvants suchas are commonly used in the art may be included. These and other suchcompounds are described in the literature, e.g., in the Merck Index,Merck & Company, Rahway, N.J. Considerations for the inclusion ofvarious components in pharmaceutical compositions are described, e.g.,in Gilman et al. (Eds.) (1990); Goodman and Gilman's: ThePharmacological Basis of Therapeutics, 8th Ed., Pergamon Press.

In yet another aspect, the invention provides a method of suppressinggrowth of a microbe, e.g., a bacterium, expressing an efflux pump, e.g.,a non-tetracycline-specific efflux pump. As illustrated by the casewhere the microbe is a bacterium, the method involves contacting thatbacterium with an efflux pump inhibitor, e.g., anon-tetracycline-specific efflux pump inhibitor, in the presence of aconcentration of antibacterial agent below the MIC of the bacterium.This method is useful, for example, to prevent or cure contamination ofa cell culture by a bacterium possessing an efflux pump. However, itapplies to any situation where such growth suppression is desirable.

In a related aspect, the invention provides a method of suppressinggrowth of a microbe, e.g., a bacterium, which involves contacting themicrobe with an efflux pump inhibitor which reduces the expression of acomponent of an efflux pump. Such an inhibitor can act on the regulationof that expression in number of different ways. It may, for example,enhance the production of a repressor molecule which prevents expressionof an efflux pump component. Another possible mechanism is if theinhibitor blocks the release of a repressor molecule. Examples of such arepressor is MarR in E. coli (Seoane and Levy, 1994, Abstr. of the Am.Soc. for Microbiol. Gen. Meeting, Las Vegas, Nev., Abstr. H-26). Anexample of a positive regulator is BmrR in Bacillus subtilis (Ahmed etal., 1994, J. Biol. Chem.).

In another related aspect, the invention provides a method for reducinga population of a microbial, e.g., a bacterial strain, involvingcontacting the population with an efflux pump inhibitor which inhibits acomponent of an efflux pump expressed in the microbe in that population,which is essential for the growth of the microbe expressing that effluxpump. In particular embodiments, that component is a cytoplasmicmembrane component. As indicated above, such efflux pump inhibitors mayact in various ways, including, but not limited to, acting directly onthe essential component, or acting to inhibit the expression of thatcomponent.

The term "reducing a population" means that the microbes of thatpopulation are being killed. This is distinguished from the action of astatic agent, e.g., a bacteriostatic agent, which prevents the bacteriafrom growing and multiplying but does not kill the microbes.Accordingly, in the context of this aspect, an "essential component" ofan efflux pump is one which is essential to the in vivo survival of themicrobe, i.e., the survival in a host.

In yet another aspect, this invention provides a method for enhancinggrowth of an animal by administering an efflux pump inhibitor to theanimal, which inhibits an efflux pump expressed in a bacterial strain inthe animal, and which inhibits the growth of that bacterial strain. Sucha growth enhancing effect may result from the reduced energy consumptionby the bacteria, which increases the food energy available to theanimal. This method is appropriate, for example, for use with cattle,swine, and fowl such as chickens and turkeys.

In an additional aspect, the invention provides novel compounds havingefflux pump activity. These compounds have chemical structures asdescribed above.

Other features and advantages of the invention will be apparent from thefollowing description of the preferred embodiments, and from the claims.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

Identification of Efflux Pump Inhibitors

Initial identification of efflux pump inhibitors having structures asdescribed for the present invention was performed using a screeningmethod as generally described in Trias et al., EFFLUX PUMP INHIBITORS,U.S. application Ser. No. 08/427,088 and Trias et al., EFFLUX PUMPINHIBITORS, U.S. application Ser. No. 08/898,477, filed Jul. 22, 1997.In particular, the screening method based on inhibition of microbialgrowth in the presence of a subinhibitory concentration of anantibacterial agent which is normally effluxed by the test microbe and aconcentration of a test compound was used for indentifying some of theactive compounds disclosed herein. In this method, inhibition of growthof the microbe is indicative that export of the antibacterial agent isinhibited by the test compound, and that the test compound is thereforean efflux pump inhibitor. The mode of action of the test compound soidentified can then be comfirmed as inhibiting active efflux. However,other screening methods for detecting efflux pump inhibitors can also beused, specifically including the additional methods described in theabove references.

Synthesis of Derivatives of Efflux Pump Inhibitors from Screening

Exemplary compounds of the present invention were synthesized by methodsas described in the Examples below. Those skilled in the art willunderstand how to synthesize additional compounds within the scope ofthis invention based on the described syntheses and the knowledge ofthose skilled in the art of chemical synthesis.

Susceptibility Testing

Particular exemplary efflux pump inhibitor compounds within the genericdescriptions of the compounds of this invention were evaluated forpotentiation effect. The in vitro microbiological data for antibioticpotentiation is presented in Tables 1-4 below. The compounds of Tables 1and 2 correspond to Structure 2 compounds, compounds of Table 3correspond to Structure 3 compounds, compounds of Table 4 to Structure 4compounds.

Potentiation effect is observed by the reduction of the minimuminhibitory concentration of levofloxacin in the presence of theexperimental efflux pump inhibitor. The activity of efflux pumpinhibitors (EPI) in combination with fluoroquinolones, such aslevofloxacin, is assessed by the checkerboard assay (AntimicrobialCombinations. In Antibiotics in Laboratory Medicine, Ed. Victor Lorian,M. D., Fourth edition, 1996, pp 333-338) using broth microdilutionmethod performed as recommended by the NCCLS (National Committee forClinical Laboratory Standards (NCCLS). 1997. Methods for Dilution ofAntimicrobial Susceptibility Tests for Bacteria That GrowAerobically--Fourth Edition; Approved Standard. NCCLS Document M7-A4,Vol 17 No.2). The test organism used is Pseudomonas aeruginosa PAM1001.The compounds of this invention demonstrate pump inhibitory activityagainst a broad-range of P. aeruginosa over-producing singular effluxpumps (MexAB, MexCD, and MexEF) and clinical strains containing multipleefflux pumps, not limited to the Mex classification. The compoundstabulated below are representative of the described invention.

In this assay, multiple dilutions of two drugs, namely the EPI andlevofloxacin, are being tested, alone and in combination, atconcentrations equal to, above and below their respective minimalinhibitory concentrations (MICs). In the case of EPI, most of thesecompounds are devoid of intrinsic antimicrobial activity and are testedat the maximum concentration of 40 μg/ml. The MIC of levofloxacinagainst P. aeruginosa PAM1001 is 4 μg/ml.

The EPI tested are readily soluble in water and stock solutions areprepared at a final concentration of 2 mg/ml. Stock solutions arefurther diluted, according to the needs of a particular assay, inMueller Hinton Broth (MHB). Stock solution can be stored at -80° C.Quinolones are solubilized according to the instructions of themanufacturers, at a concentration of 1 mg/ml. They are then furtherdiluted in MHB. Stock solution can be stored at -80° C.

The checkerboard assay is performed in microtiter plates. Levofloxacinis diluted in the x axis, each column containing a single concentrationof levofloxacin. The EPI is diluted in the y axis, each row containingan equal concentration of EPI. The result of these manipulations is thateach well of the microtiter plate contains a unique combination ofconcentrations of the two agents. Each EPI are tested independently.

The assay is performed in MHB with a final bacterial inoculum of 5×10⁵CFU/ml (from an early-log phase culture). Microtiter plates areincubated during 20 h at 35° C. and are read using a microtiterplatereader (Molecular Devices) at 650 nm as well as visual observation usinga microtiterplate reading mirror. The MIC is defined as the lowestconcentration of quinolone, within the combination, at which the visiblegrowth of the organism is completely inhibited.

    ______________________________________                                        Efflux Pump Inhibitors (EPIs) for Table 1                                     Comp  Structure                                                               ______________________________________                                        1     Phenylalanyl-ornithine quinoline-3-amide                                2     Phenylalanyl-ornithine quinoline-8-amide                                3     Phenylalanyl-ornithine 2-methylquinoline-8-amide                        4     Alanyl-phenylalanyl-arginine 2-naphthylamide                            5     D-Alanyl-phenylalanyl-arginine 2-naphthylamide                          6     Valyl-phenylalanyl-arginine 2-naphthylamide                             7     4-Fluorophenylalanyl-ornithine quinoline-3-amide                        8     4-Fluorophenylalanyl-ornithine quinoline-8-amide                        9     4-Iodophenylalanyl-ornithine quinoline-3-amide                          10    4-Iodophenylalanyl-ornithine quinoline-8-amide                          11    Homophenylalanyl-ornithine quinoline-3-amide                            12    Homophenylalanyl-ornithine quinoline-8-amide                            13    Homophenylalanyl-ornithine quinoline-6-amide                            14    Homophenylalanyl-ornithine isoquinoline-5-amide                         15    Phenylalanyl-N.sub.α -methylarginine 2-naphthylamide              16    Phenylalanyl-N.sub.α -methylornithine 2-naphthylamide             17    Phenylalanyl-N.sub.α -methylornithine 2-(naphthylmethyl)amide     18    Phenylalanyl-N.sub.α -methylornithine 2,2-diphenylethylamide      19    4-Fluorophenylalanyl-N.sub.α -methylornithine                           2-naphthylamide                                                         20    4-Iodophenylalanyl-N.sub.α -methylornithine                             4-fluorophenethylamide                                                  21    Tyrosyl-N.sub.α -methylornithine 2-naphthylamide                  22    Homophenylalanyl-N.sub.α -methylornithine 4-fluorophenethylami          de                                                                      23    Homophenylalanyl-N.sub.α -methylornithine 4-methylphenethylami          de                                                                      24    Homophenylalanyl-N.sub.α -methylornithine 2,2-diphenylethylami          de                                                                      25    Homophenylalanyl-N.sub.α -methylornithine                               1,2,3,4-tetrahydronaphthyl-5-amide                                      26    Homophenylalanyl-N.sub.α -methylornithine 3-phenylpropylamide     27    Homophenylalanyl-N.sub.α -methylornithine 3-(4-                         methylphenyl)propylamide                                                28    Homophenylalanyl-N.sub.α -methylornithine 3-(4-                         methoxyphenyl)propylamide                                               29    Homophenylalanyl-N.sub.α -methylornithine 3-(4-                         fluorophenyl)propylamide                                                30    β-(2-Thiaizolyl)alanyl-N.sub.α -methylornithine                    2-naphthylamide                                                         31    4-(Dimethylaminoethoxy)phenylalanyl-N.sub.α -methylornithine            2-                                                                            naphthylamide                                                           32    4-(O-Methylcarboxyamido)phenylalanyl-N.sub.α -methylornithine           2-                                                                            naphthylamide                                                           33    β-(1-Naphthyl)alanyl-N.sub.α -methylornithine                      benzylamide                                                             34    β-(2-Naphthyl)alanyl-N.sub.α -methylornithine                      benzylamide                                                             35    β-(2-Naphthyl)alanyl-N.sub.α -methylornithine                      4-hydroxyphenethylamide                                                 36    Leucyl-N.sub.α -methylornithine 2-naphthylamide                   37    β-(Cyclohexyl)alanyl-N.sub.α -methylornithine                      phenethylamide                                                          38    Glycyl-N.sub.α -methylornithine 2-(cyclohexyl)ethylamide          39    Glycyl-N.sub.α -(phenethyl)ornithine 2-naphthylamide              40    Glycyl-N.sub.α -(phenethyl)ornithine 3-phenylpropylamide          41    Glycyl-N.sub.α -(phenethyl)ornithine quinoline-3-amide            42    Glycyl-N.sub.α -(phenethyl)ornithine 5-indanylamide               43    Glycyl-N.sub.α -(2-hydroxyphenethyl)ornithine                           3-phenylpropylamide                                                     44    Glycyl-N.sub.α -(3-phenylpropyI)ornithine 3-phenylpropylamide     45    Glycyl-N.sub.α -(isoamyl)ornithine 3-phenylpropylamide            46    Glycyl-N.sub.α -(2-benzoxazolylmethyl)ornithine                         3-phenylpropylamide                                                     47    Glycyl-N.sub.α -(3-quinolinylmethyl)ornithine                           3-phenylpropylamide                                                     48    βAlanyl-N.sub.α -(phenethyl)ornithine 3-phenylpropylamide    49    Acetimidoylglycyl-N.sub.α -(phenethyl)ornithine                         3-phenylpropylamide                                                     50    Glycyl-N.sub.α -(phenethyl)lysine 3-phenylpropylamide             51    β-Alanyl-N.sub.α -(phenethyl)lysine 3-phenylpropylamide      52    4-Aminobutyryl-N.sub.α -(phenethyl)diaminopropionic acid 3-             phenylpropylamide                                                       53    4-Aminobutyryl-N.sub.α -(phenethyl)diaminopropionic acid                quinoline-2-amide                                                       54    Glycyl-N.sub.α -(phenethyl)diaminobutyric acid                          3-phenylpropylamide                                                     55    β-Alanyl-N.sub.α -(phenethyl)diaminobutyric acid                   3-phenylpropylamide                                                     56    4-Aminobutyryl-N.sub.α -(phenethyl)diaminobutyric acid 3-               phenylpropylamide                                                       ______________________________________                                    

    ______________________________________                                        Efflux Pump Inhibitors (EPIs) for Table 2                                     Comp  Structure                                                               ______________________________________                                        1     D-Arginyl-D-phenylalanine quinoline-3-amide                             2     D-Ornithyl-D-phenylalanine 2,2-diphenylethylamide                       3     D-Ornithyl-D-phenylalanine 2-naphthylamide                              4     Ornithyl-phenylalanine 1,2,3,4-tetrahydronaphthyl-5-amide               5     D-Ornithyl-D-phenylalanine 1,2,3,4-tetrahydronaphthyl-5-amide           6     Ornithyl-phenylalanine quinoline-3-amide                                7     D-Ornithyl-D-phenylalanine quinoline-3-amide                            8     Ornithyl-phenylalanine quinoline-8-amide                                9     D-Ornithyl-D-phenylalanine quinoline-8-amide                            10    D-Ornithyl-D-phenylalanine 3-phenylpropylamide                          11    D-Ornithyl-D-4-methylphenylalanine 2-naphthylamide                      12    D-Ornithyl-D-(N-methyl)phenylalanine 2-naphthylamide                    13    D-Lysyl-D-phenylalanine 2-naphthylamide                                 14    D-Ornithyl-D-homophenylalanine quinoline-3-amide                        15    D-Ornithyl-D-homophenylalanine 2-naphthylamide                          16    D-Ornithyl-D-homophenylalanine quinoline-8-amide                        17    D-Ornithyl-D-homophenylalanine 2,2-diphenylethylamide                   18    D-Ornithyl-homophenylalanine quinoline-3-amide                          19    Ornithyl-D-homophenylalanine quinoline-3-amide                          20    D-Ornithyl-D-homophenylalanine quinoline-3-amide                        21    D-Ornithyl-D-homophenylalanine quinoline-8-amide                        22    D-Ornithyl-D-homophenylalanine (2-quinolinylmethyl)amide                23    D-Ornithyl-D-homophenylalanine (3-quinolinylmethyl)amide                24    D-Ornithyl-D-homophenylalanine 1-fluoronaphthyl-2-amide                 25    D-Ornithyl-D-homophenylalanine 2-naphthylamide                          26    D-Ornithyl-D-homophenylalanine 3-phenylpropylamide                      27    D-Ornithyl-D-homophenylalanine 4-methylphenethylamide                   28    D-Ornithyl-D-homophenylalanine 4-fluorophenethylamide                   29    D-Lysyl-D-homophenylalanine 2-naphthylamide                             30    D-Ornithyl-D-β-(2-naphthyl)alanine benzylamide                     31    D-Ornithyl-D-β-(1-naphthyl)alanine benzylamide                     32    D-Ornithyl-D-β-(2-naphthyl)alanine 4-hydroxyphenethylamide         33    D-Ornithyl-D-β-(2-naphthyl)alanine iso-amylamide                   34    D-Ornithyl-D-β-(2-naphthyl)alanine 2-hydroxybenzylamide            35    D-Ornithyl-D-β-(2-naphthyl)alanine phenethylamide                  36    D-Ornithyl-D-β-(3-quinolinyl)alanine 3,3-dimethylbutylamide        37    D-Ornithyl-D-β-(3-quinolinyl)alanine 4-(t-butyl)phenylamide        38    D-Ornithyl-D-β-(3-quinolinyl)alanine 4-methylphenethylamide        39    D-Ornithyl-D-β-(3-quinolinyl)alanine 4-ethylbenzylamide            40    D-Ornithyl-D-β-(3-quinolinyl)alanine 3-phenylpropylamide           41    D-Ornithyl-D-β-(3-quinolinyl)alanine 2,3-trimethylenepyridyl-5-          amide                                                                   42    D-N.sub.α -(C-Amidino)arginyl-D-β-(2-naphthyl)alanine              benzylamide                                                             43    D-Ornithyl-D-leucine 4-fluorophenethylamide                             44    D-Ornithyl-D-leucine 3-phenylpropylamide                                45    D-Ornithyl-D-valine 2-naphthylamide                                     46    D-Ornithyl-D-β-(t-butyl)alanine quinoline-3-amide                  47    D-Diaminobutyryl-D-homophenylalanine quinoline-3-amide                  48    D-Lysyl-D-β-(t-butyl)alanine quinoline-3-amide                     49    D-Lysyl-D-homophenylalanine quinoline-3-amide                           50    D-Lysyl-D-homophenylalanine (1-isoquinolinylmethyl)amide                51    D-Lysyl-D-homophenylalanine (2-quinolinylmethyl)amide                   52    D-Lysyl-D-homophenylalanine (3-quinolinylmethyl)amide                   53    D-Lysyl-D-β-(3-quinolinyl)alanine 4-ethylbenzylamide               ______________________________________                                    

    ______________________________________                                        Efflux Pump Inhibitors (EPIs) for Table 3                                     Comp  Structure                                                               ______________________________________                                        1     D-Ornithyl-N-(benzyl)glycine 2-naphthylamide                            2     D-Ornithyl-N-(benzyl)glycine 3-phenylpropylamide                        3     D-Ornithyl-N-(phenethyl)glycine 2-naphthylamide                         4     D-Ornithyl-N-(phenethyl)glycine 3-phenylpropylamide                     5     Ornithyl-N-(phenethyl)glycine 3-phenylpropylamide                       6     D-Ornithyl-N-(phenylpropyl)glycine 3-phenylpropylamide                  7     D-Ornithyl-β-(N-isopropyl)alanine 2-naphthylamide                  8     D-Ornithyl-β-(N-isopropyl)alanine quinoline-3-amide                9     D-Ornithyl-β-(N-isoamyl)alanine 2-naphthylamide                    10    D-Ornithyl-β-(N-isoamyl)alanine quinoline-3-amide                  11    D-Ornithyl-β-(N-benzyl)alanine 2-naphthylamide                     12    D-Ornithyl-β-(N-benzyl)alanine 3-phenylpropylamide                 13    D-Ornithyl-β-(N-benzyl)alanine quinoline-3-amide                   14    D-Ornithyl-β-(N-phenethyl)alanine quinoline-3-amide                15    D-Ornithyl-β-(N-phenethyl)alanine 2-naphthylamide                  16    D-Ornithyl-β-(N-phenethyl)alanine 3-phenylpropylamide              17    D-Ornithyl-β-(N-cyclohexylmethyl)alanine 2-naphthylamide           18    D-Ornithyl-β-(N-cyclohexylmethyl)alanine quinoline-3-amide         19    D-Ornithyl-β-(N-cyclohexylmethyl)alanine                                 3-phenylpropylamide                                                     20    D-Ornithyl-β-(N-phenylpropyl)alanine 2-naphthylamide               21    Omithyl-β-(N-phenylpropyl)alanine 2-naphthylamide                  22    D-Ornithyl-β-(N-phenylpropyl)alanine quinoline-3-amide             23    Ornithyl-β-(N-phenylpropyl)alanine quinoline-3-amide               24    Ornithyl-β-(N-phenylpropyl)alanine 3-phenylpropylamide             25    D-Ornithyl-β-(N-phenylpropyl)alanine                                     (cyclohexylmethyl)amide                                                 26    D-Ornithyl-β-[N-(4-methylphenyl)propyl]alanine                           2-naphthylamide                                                         27    D-Ornithyl-β-[N-(4-methylphenyl)propyl]alanine                           quinoline-3-amide                                                       28    D-Ornithyl-β-(N-(4-methoxyphenethyl)alanine 2-naphthylamide        29    D-Ornithyl-β-(N-4-methoxyphenethyl)alanine quinoline-3-amide       30    D-Ornithyl-β-(N-4-methylphenethyl)alanine 2-naphthylamide          31    D-Ornithyl-β-(N-4-methylphenethyl)alanine 1-fluoronaphthyl-2-            amide                                                                   32    D-Ornithyl-β-(N-4-methylphenethyl)alanine quinoline-3-amide        33    D-Ornithyl-β-(N-4-fluorophenylpropyl)alanine 2-naphthylamide       34    D-Ornithyl-β-(N-4-fluorophenylpropyl)alanine                             quinolinyl-3-amide                                                      35    D-Ornithyl-β-(N-cyclopropylmethyl)alanine 2-naphthylamide          36    D-Ornithyl-β-(N-cyclopropylmethyl)alanine quinolinyl-3-amide       37    D-Ornithyl-β-[N-(3,3-dimethylbutyl)]alanine 2-naphthylamide        38    D-Ornithyl-β-[N-(3,3-dimethylbutyl)]alanine quinolinyl-3-amide     39    D-Ornithyl-β-[N-(isobutyl)]alanine 2-naphthylamide                 40    D-Ornithyl-β-[N-(isobutyl)]alanine quinoline-3-amide               41    D-Ornithyl-β-[N-(3-ethoxypropyl)]alanine 2-naphthylamide           42    D-Ornithyl-β-[N-(ethylthioethyl)]alanine 2-naphthylamide           43    D-Ornithyl-β-[N-(ethylthioethyl)]alanine quinoline-3-amide         ______________________________________                                    

    ______________________________________                                        Efflux Pump Inhibitors (EPIs) for Table 4                                     Comp  Structure                                                               ______________________________________                                        1     Phenylalanyl-ornithinol 2-naphthyl ether                                2     Phenylalanyl-ornithinthiol 2-naphthyl thioether                         3     Homophenylalanyl-ornithinol 2-naphthyl ether                            4     Homophenylalanyl-ornithinthiol 2-benzothiazole thioether                5     β-(2-Naphthyl)alanyl-ornithinthiol 2-benzothiazole thioether       6     Homophenylalanyl-N.sub.α -methylornithinol 2-naphthyl ether       7     Homophenylalanyl-N.sub.α -methylornithinol 2-benzothiazole              thioether                                                               8     D-Phenylalanyl-N.sub.α -methylornithinol 2-benzothiazole                thioether                                                               10    Phenylalanyl-N.sub.α -methylornithinol 2-benzothiazole                  thioether                                                               11    Homophenylalanyl-N.sub.α -methylargininol 2-naphthyl ether        12    D-Ornithyl-D-phenylalaninol 2-naphthyl ether                            13    D-Lysyl-D-phenylalaninol 2-naphthyl ether                               14    Ornithyl-N.sub.α -methylphenylalaninol 2-naphthyl ether           15    Ornithyl-phenylalaninthiol 2-benzothiazole thioether                    16    D-Ornithyl-D-phenylalaninthiol 2-benzothiazole thioether                17    O-Benzylseryl-N.sub.α -methylornithinol 2-naphthyl ether          18    N.sub.α -(C-Amidino)homophenylalanyl-N.sub.α -methylargi          ninol                                                                         2-naphthyl ether                                                        19    D-Ornithyl-D-phenylalaninol 2-quinolinyl ether                          20    D-Ornithyl-D-phenylalaninol 8-quinolinyl ether                          21    D-Lysyl-D-phenylalaninthiol 2-benzothiazolyl thioether                  22    D-Ornithyl-D-valinol 2-naphthyl ether                                   23    D-Ornithyl-D-valinol 2-quinolinyl ether                                 24    D-Ornithyl-D-phenylalaninthiol 2-naphthyl thioether                     25    D-Ornithyl-D-phenylalaninthiol 3-quinolinyl thioether                   26    D-Ornithyl-D-leucinethiol 2-naphthyl thioether                          27    D-Ornithyl-D-leucinethiol 2-quinolinyl thioether                        28    D-Lysyl-D-phenylalaninethiol 3-quinolinyl thioether                     29    D-Lysyl-D-phenylalaninethiol 2-naphthyl thioether                       30    N-Ornithyl-N-benzylaminoethanol 2-naphthyl ether                        31    D-Ornithyl-N-benzylaminoethanol 2-naphthyl ether                        32    Tyrosyl-N.sub.α -methylornithinol 2-naphthyl ether                33    Homophenylalanyl-N-(3-aminopropyl)aminoethanol 2-naphthyl                     ether                                                                   34    D-Ornithyl-N-(phenethyl)aminoethanol 2-naphthyl ether                   35    Ornithyl-N-(phenethyl)aminoethanol 2-naphthyl ether                     36    β-(Cyclohexyl)alanine N-(3-aminopropyl)-3-                               (cyclohexyl)propylamide                                                 37    D-Ornithyl-N-(phenethyl)aminopropanol 2-quinolinyl ether                38    D-Ornithyl-N-(benzyl)aminopropanol 2-quinolinyl ether                   39    D-Ornithyl-N-(phenethyl)aminoethanethiol 3-phenylpropyl                       thioether                                                               40    D-Ornithine N-(isoamylaminoethyl)phenylpropylamide                      41    D-Ornithyl-D-phenylalaninethiol benzyl thioether                        42    Homophenylalanyl-ornithinethiol 2-phenethyl thioether                   43    Homophenylalanine N-(3-aminopropyl)-3-phenylpropylamide                 44    D-Ornithyl-N-(phenethyl)aminoethanethiol S-benzyl thioether             45    D-Ornithyl-N-(phenethyl)aminoethanethiol S-(4-ethylbenzyl)                    thioether                                                               ______________________________________                                    

                                      TABLE 1                                     __________________________________________________________________________    Levofloxacin MIC Against P. aeruginosa PAM1001 in Presence of Efflux Pump     Inhibitor (EPI)                                                               Minimum Inhibitory Concentration (μg/ml)                                          EPI Conc. 0                                                                          EPI Conc.                                                                            EPI Conc.                                                                            EPI Conc.                                                                           EPI Conc.                                                                           EPI Conc. 10                                                                          EPI Conc.                                                                             EPI Conc. 40          Compound                                                                             μg/ml                                                                             0.625 μg/ml                                                                       1.25 μg/ml                                                                        2.5 μg/ml                                                                        5 μg/ml                                                                          μg/ml                                                                              μg/ml                                                                              μg/ml              __________________________________________________________________________    1      4      4      4      4     4     2       0.50    0.06                  2      4      4      4      4     4     1       0.50    0.06                  3      4      4      4      4     4     2       1       0.50                  4      4      4      4      4     4     0.06    0.03    0.03                  5      4      4      4      4     4     0.25    0.03    0.03                  6      4      4      4      4     4     2       0.015   0.008                 7      4      4      4      4     2     1       0.50    0.06                  8      4      4      4      4     2     1       0.50    0.25                  9      4      4      4      1     0.06  0.06    0.06    0.06                  10     4      4      4      2     0.50  0.25    0.25    0.125                 11     4      4      4      2     1     0.06    0.03    0.03                  12     4      4      4      2     0.50  0.125   0.125   0.125                 13     4      4      4      4     2     1       0.50    0.06                  14     4      4      4      4     4     2       1       0.25                  15     4      4      4      4     1     0.06    0.03    0.015                 16     4      4      4      4     0.50  0.03    0.015   0.015                 17     4      4      4      4     4     1       0.06    0.03                  18     4      4      4      4     4     2       0.50    0.25                  19     4      4      4      4     0.06  0.06    0.03    0.015                 20     4      4      4      2     0.06  0.03    0.03    0.03                  21     4      4      2      1     0.25  0.125   0.03    0.008                 22     4      4      4      4     2     1       0.125   0.125                 23     4      4      4      2     1     0.25    0.25    0.25                  24     4      4      4      4     4     1       0.06    0.008                 25     4      4      4      4     1     0.06    0.06    0.06                  26     4      4      4      2     1     0.125   0.125   0.06                  27     4      4      4      1     0.125 0.125   0.125   0.06                  28     4      4      4      4     2     1       0.06    0.125                 29     4      4      4      2     1     0.125   0.125   0.125                 30     4      4      4      4     4     2       0.50    0.03                  31     4      4      4      2     1     1       0.50    0.25                  32     4      4      4      4     2     1       0.50    0.06                  33     4      4      4      4     4     2       1       0.50                  34     4      4      4      4     4     2       0.125   0.125                 35     4      4      4      4     2     1       0.50    0.125                 36     4      4      2      2     0.125 0.06    0.015   0.015                 37     4      4      4      4     1     0.25    0.125   0.125                 38     4      4      4      4     4     2       0.50    0.50                  39     4      2      0.50   0.06  0.03  0.03    0.03    0.03                  40     4      4      1      0.50  0.25  0.125   0.06    0.125                 41     4      4      4      1     1     0.25    0.25    0.25                  42     4      2      1      0.03  0.015 0.015   0.015   0.015                 43     4      4      4      2     1     0.25    0.25    1                     44     4      4      2      1     0.50  0.25    0.25    0.25                  45     4      4      4      2     1     0.50    0.50    0.125                 46     4      4      4      1     0.25  0.125   0.125   0.250                 47     4      4      4      2     1     0.25    0.25    0.125                 48     4      4      4      1     1     0.25    0.25    0.25                  49     4      4      4      4     2     1       0.25    0.25                  50     4      4      4      1     0.50  0.125   0.06    0.06                  51     4      4      4      2     1     0.125   0.125   0.125                 52     4      4      4      4     1     0.50    0.125   0.125                 53     4      4      4      4     1     0.25    0.06    0.03                  54     4      2      2      1     0.25  0.25    0.06    0.06                  55     4      4      2      2     0.50  0.25    0.125   0.125                 56     4      4      4      2     0.50  0.25    0.25    0.06                  __________________________________________________________________________

                                      TABLE 2                                     __________________________________________________________________________    Levofloxacin MIC Against P. aeruginosa PAM1001 in Presence of Efflux Pump     Inhibitor (EPI)                                                               Minimum Inhibitory Concentration (μg/ml)                                          EPI Conc. 0                                                                          EPI Conc.                                                                            EPI Conc.                                                                            EPI Conc.                                                                           EPI Conc.                                                                           EPI Conc. 10                                                                          EPI Conc.                                                                             EPI Conc. 40          Compound                                                                             μg/ml                                                                             0.625 μg/ml                                                                       1.25 μg/ml                                                                        2.5 μg/ml                                                                        5 μg/ml                                                                          μg/ml                                                                              μg/ml                                                                              μg/ml              __________________________________________________________________________    1      4      4      4      4     2     0.06    0.0135  0.015                 2      4      4      4      4     4     4       1       0.25                  3      4      4      4      4     4     1       0.06    0.03                  4      4      4      4      4     4     2       0.50    0.125                 5      4      4      4      4     4     1       0.25    0.125                 6      4      4      4      4     2     2       0.50    0.125                 7      4      4      4      4     4     4       0.50    0.125                 8      4      4      4      4     2     2       0.50    0.06                  9      4      4      4      4     4     2       1       0.06                  10     4      4      4      4     4     2       0.25    0.25                  11     4      4      4      2     0.125 0.06    0.03    NA                    12     4      4      4      4     2     0.50    0.06    0.03                  13     4      4      4      4     2     1       0.125   0.06                  14     4      4      4      4     2     1       0.50    0.03                  15     4      4      4      2     1     0.06    0.06    0.06                  16     4      4      4      4     2     1       0.125   0.03                  17     4      4      4      4     2     0.50    0.25    0.03                  18     4      4      4      4     4     2       0.125   0.125                 19     4      4      4      4     4     0.50    0.06    0.015                 20     4      4      4      2     1     0.125   0.03    0.015                 21     4      4      4      4     2     0.50    0.03    0.008                 22     4      4      4      4     1     0.25    0.06    0.03                  23     4      4      4      4     2     0.50    0.50    0.06                  24     4      4      4      2     0.25  0.03    0.03    0.06                  25     4      4      4      2     1     0.06    0.06    0.06                  26     4      4      4      4     2     0.125   0.125   0.06                  27     4      4      4      2     2     2       2       2                     28     4      4      4      4     1     0.50    0.25    0.125                 29     4      4      4      2     0.125 0.06    0.06    0.06                  30     4      4      4      0.50  0.25  0.03    0.03    0.03                  31     4      4      4      4     2     0.125   0.03    0.03                  32     4      4      4      4     2     0.50    0.125   0.06                  33     4      4      4      2     0.25  0.06    0.06    0.06                  34     4      4      4      2     0.50  0.25    0.06    0.50                  35     4      4      4      4     2     0.25    0.125   0.06                  36     4      4      4      4     2     1       0.25    0.50                  37     4      4      4      4     2     0.008   0.015   0.008                 38     4      4      4      4     4     1       0.125   0.03                  39     4      4      4      4     2     0.25    0.03    0.25                  40     4      4      4      4     4     2       0.50    0.03                  41     4      4      4      2     4     0.015   0.015   0.03                  42     4      2      2      0.50  0.125 0.25    0.25    0.25                  43     4      4      4      4     4     2       2       0.50                  44     4      4      4      4     4     1       0.50    0.25                  45     4      4      4      4     4     2       1       0.50                  46     4      4      4      4     2     1       0.50    0.25                  47     4      4      4      4     2     1       0.125   0.03                  48     4      4      4      4     2     1       0.50    0.25                  49     4      4      2      2     0.50  0.06    0.03    0.03                  50     4      4      4      4     2     0.50    0.25    0.03                  51     4      4      4      4     4     1       0.50    0.06                  52     4      4      4      4     2     1       0.25    0.03                  53     4      4      4      4     4     0.06    0.015   0.06                  __________________________________________________________________________

                                      TABLE 3                                     __________________________________________________________________________    Levofloxacin MIC Against P. aeruginosa PAM1001 in Presence of Efflux Pump     Inhibitor (EPI)                                                               Minimum Inhibitory Concentration (μg/ml)                                          EPI Conc. 0                                                                          EPI Conc.                                                                            EPI Conc.                                                                            EPI Conc.                                                                           EPI Conc.                                                                           EPI Conc. 10                                                                          EPI Conc.                                                                             EPI Conc. 40          Compound                                                                             μg/ml                                                                             0.625 μg/ml                                                                       1.25 μg/ml                                                                        2.5 μg/ml                                                                        5 μg/ml                                                                          μg/ml                                                                              μg/ml                                                                              μg/ml              __________________________________________________________________________    1      4      4      4      4     4     1       0.25    0.06                  2      4      4      4      4     4     2       1       0.125                 3      4      4      4      4     2     1       0.25    0.03                  4      4      4      4      4     2     0.25    0.125   0.125                 5      4      4      4      4     2     1       0.25    0.125                 6      4      4      4      2     1     0.06    0.06    0.06                  7      4      4      4      4     2     0.50    0.25    0.06                  8      4      4      4      4     4     2       2       0.50                  9      4      4      4      2     0.125 0.06    0.06    0.06                  10     4      4      4      4     2     0.50    0.125   0.03                  11     4      4      4      1     0.25  0.06    0.008   0.008                 12     4      4      4      4     2     1       0.50    0.125                 13     4      4      4      2     1     0.50    0.25    0.015                 14     4      4      4      4     2     0.50    0.03    0.015                 15     4      4      4      2     0.06  0.015   0.015   0.015                 16     4      4      4      4     4     2       0.50    0.06                  17     4      4      2      2     0.25  0.06    0.06    0.06                  18     4      4      4      2     1     0.50    0.03    0.03                  19     4      4      4      4     2     0.50    0.125   0.125                 20     4      4      2      1     0.03  0.03    0.03    0.03                  21     4      4      2      1     0.03  0.03    0.06    0.03                  22     4      4      4      2     1     0.125   0.03    0.06                  23     4      4      4      2     0.25  0.06    0.03    0.015                 24     4      4      4      4     2     0.25    0.125   0.125                 25     4      4      4      4     2     1       0.50    0.25                  26     4      4      4      2     0.06  0.015   0.03    NA                    27     4      4      4      1     0.06  0.03    0.03    0.03                  28     4      4      4      2     0.015 0.008   0.015   0.015                 29     4      4      4      4     2     0.25    0.03    0.008                 30     4      4      4      2     0.25  0.015   0.015   0.015                 31     4      4      2      0.125 0.03  0.03    0.06    NA                    32     4      4      2      2     0.125 0.008   0.008   0.015                 33     4      4      2      0.06  0.015 0.015   0.03    NA                    34     4      4      4      2     1     0.25    0.015   0.03                  35     4      4      4      4     2     1       0.25    0.06                  36     4      4      4      4     4     2       1       0.50                  37     4      4      4      1     0.25  0.03    0.03    0.06                  38     4      4      4      4     1     0.25    0.06    0.015                 39     4      4      4      4     2     0.50    0.125   0.03                  40     4      4      4      4     4     2       1       0.25                  41     4      4      4      4     4     1       0.50    0.125                 42     4      4      4      4     2     0.50    0.06    0.015                 43     4      4      4      4     4     2       0.50    0.125                 __________________________________________________________________________

                                      TABLE 4                                     __________________________________________________________________________    Levofloxacin MIC Against P. aeruginosa PAM1001 in Presence of Efflux Pump     Inhibitor (EPI)                                                               Minimum Inhibitory Concentration (μg/ml)                                          EPI Conc. 0                                                                          EPI Conc.                                                                            EPI Conc.                                                                            EPI Conc.                                                                           EPI Conc.                                                                           EPI Conc. 10                                                                          EPI Conc.                                                                             EPI Conc. 40          Compound                                                                             μg/ml                                                                             0.625 μg/ml                                                                       1.25 μg/ml                                                                        2.5 μg/ml                                                                        5 μg/ml                                                                          μg/ml                                                                              μg/ml                                                                              μg/ml              __________________________________________________________________________    1      4      4      4      2     1     0.03    0.03    0.03                  2      4      4      4      4     2     1       0.50    0.06                  3      4      4      4      2     0.015 0.015   0.03    NA                    4      4      4      4      0.25  0.03  0.03    0.06    0.06                  5      4      4      4      4     2     0.125   0.06    0.06                  6      4      4      4      4     0.03  0.03    0.03    0.008                 7      4      4      4      2     0.03  0.06    0.06    0.03                  8      4      4      4      4     4     4       2       0.50                  9      4      4      4      4     1     1       0.06    0.125                 10     4      4      4      2     0.125 0.03    NA      NA                    11     4      4      2      1     0.125 0.03    0.03    0.03                  12     4      4      4      2     1     0.06    0.03    0.06                  13     4      4      4      2     1     0.125   0.125   0.125                 14     4      4      4      4     2     2       2       1                     15     4      4      4      2     1     0.25    0.06    0.06                  16     4      4      4      4     2     0.015   0.03    0.015                 17     4      4      4      4     0.50  0.50    NA      NA                    18     4      4      4      4     2     0.50    0.125   0.25                  19     4      4      4      4     4     2       1       0.125                 20     4      4      4      2     1     0.06    0.06    0.125                 21     4      4      4      4     2     1       0.25    0.06                  22     4      4      4      4     4     2       1       0.50                  23     4      4      4      2     1     0.06    0.06    0.06                  24     4      4      4      4     2     1       0.125   0.03                  25     4      4      2      0.25  0.125 0.06    0.125   0.125                 26     4      4      4      4     1     0.25    0.125   0.03                  27     4      4      4      4     2     2       1       1                     28     4      4      4      2     2     0.25    0.125   0.125                 29     4      4      2      0.25  0.06  0.015   0.015   0.015                 30     4      4      2      0.25  0.03  0.03    0.03    0.015                 31     4      4      4      4     0.50  0.125   0.015   0.015                 32     4      4      2      0.25  0.125 0.125   0.06    NA                    33     4      4      4      2     0.06  0.03    0.03    0.03                  34     4      4      4      2     0.06  0.06    0.06    0.03                  35     4      4      4      2     0.25  0.25    NA      NA                    36     4      4      4      4     2     0.50    0.50    NA                    37     4      4      4      4     2     0.06    0.03    0.06                  38     4      4      4      4     1     0.25    0.125   0.03                  39     4      4      4      4     4     2       0.50    0.25                  40     4      4      4      0.50  0.50  0.125   0.125   0.125                 41     4      2      0.25   0.06  0.125 0.06    0.06    NA                    42     4      4      4      4     1     0.50    0.50    0.50                  43     4      4      4      4     2     0.50    0.25    0.125                 44     4      4      4      4     1     0.25    0.125   0.06                  __________________________________________________________________________

In vivo Evaluation of Efflux Pump Inhibitor Compounds

Inhibitors of the bacterial efflux pumps are generally initiallycharacterized in vitro. Those which show effective inhibition of thepump(s) and which show synergistic activity with antibiotics areselected for evaluation in vivo. Efficacy testing will be done usingstandard procedures. Primary efficacy evaluation may be done using themurine septicemia model (M. G. Bergeron, 1978, Scand. J. Infect. Dis.Suppl. 14:189-206; S. D. Davis, 1975, Antimicrob. Agents Chemother.8:50-53). In this model a supra-lethal dose of bacteria is used tochallenge the rodents. Treatment is initiated, varying either or bothtime(s) of treatment and dose of antibiotic. In these experiments boththe antibiotic and the efflux pump inhibitor doses are varied. Apositive result is indicated by significant increase in protection fromthe lethal infection by the combination of the potentiator (the effluxpump inhibitor) and the antibiotic versus the antibiotic alone.

A second efficacy model which is used is the mouse soft tissue infectionmodel (Vogelman et al., 1988, J. Infect. Dis. 157:287-298). In thismodel anesthetized mice are infected with an appropriate titer ofbacteria in the muscle of the hind thigh. Mice are either neutropenic(cyclophosphamide treated at 125 mg/kg on days -4,-2, and 0) orimmunocompetent. The infecting dose is commonly 10⁵ -10⁶ colony formingunits per animal. Treatment with the combination of the efflux pumpinhibitor and/or antibiotics follows infection, or can occur beforeinfection. The proliferation (or death) of the bacteria within the thighmuscle is monitored over time. Effective combinations show greateractivity than the antibiotic alone. Activity is defined as reduction ingrowth rate of the test bacteria in the murine tissue.

Another model useful for assessing the effectiveness of the efflux pumpinhibitors is the diffusion chamber model (Malouin et al., 1990, Infect.Immun. 58:1247-1253; Day et al., J. Infect. 2:39-51; Kelly et al., 1989,Infect. Immun. 57:344-350). In this model rodents have a diffusionchamber surgically placed in their peritoneal cavity. The chamber canconsist of a polypropylene cylinder with semipermeable membranescovering the cylinder ends. Diffusion of peritoneal fluid into and outof the chamber provides nutrients for the microbes. The proliferation ofthe bacteria in the presence and absence of the antibiotic/efflux pumpinhibitor is compared to the antibiotic alone. Dose ranging of thecombination and the antibiotic alone are done to assess effectiveness ofthe antimicrobial and combinations.

A tertiary model useful as a stringent test of the efflux pumpinhibitor/antibiotic combination is the endocarditis model (J. Santoroand M. E. Levinson, 1978, Infect. Immun. 19:915-918). Either rats orrabbits are effectively used in this model. The effectiveness ofcombinations of efflux inhibitor and antibiotic are compared toantibiotic alone. The end point is usually viable cells remaining in thecardiac vegetations at the end of treatment.

The examples of infection models provided are not limiting. Asunderstood by those skilled in the art, other models can be utilized asappropriate for a specific infecting microbe. In particular, cell-basedinfection models may be used in some circumstances instead of animalmodels.

Pharmaceutical Compositions and Modes of Administration

The particular compound that is an efflux pump inhibitor can beadministered to a patient either by itself, or in combination with anantimicrobial, e.g., antibacterial, agent, or in pharmaceuticalcompositions where it is mixed with suitable carriers or excipient(s). Acombination of an efflux pump inhibitor with an antimicrobial agent canbe of at least two different types. In one, a quantity of an efflux pumpinhibitor is combined with a quantity of an antimicrobial agent in amixture, e.g., in a solution or powder mixture. In such mixtures, therelative quantities of the inhibitor and the antimicrobial agent may bevaried as appropriate for the specific combination and expectedtreatment. In a second type of combination an inhibitor and anantimicrobial agent can be covalently linked in such manner that thelinked molecule can be cleaved within the cell. However, the term "incombination" can also refer to other possibilities, including serialadministration of an inhibitor and another antimicrobial agent. Inaddition, an efflux pump inhibitor and/or another antimicrobial agentmay be administered in pro-drug forms, i.e. the compound is administeredin a form which is modified within the cell to produce the functionalform. In treating a patient exhibiting a disorder of interest, atherapeutically effective amount of an agent or agents such as these isadministered. A therapeutically effective dose refers to that amount ofthe compound(s) that results in amelioration of symptoms or aprolongation of survival in a patient, and may include elimination of amicrobial infection.

Toxicity and therapeutic efficacy of such compounds can be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., for determining the LD₅₀ (the dose lethal to 50% of thepopulation) and the ED₅₀ (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio LD₅₀ /ED₅₀.Compounds which exhibit large therapeutic indices are preferred. Thedata obtained from these cell culture assays and animal studies can beused in formulating a range of dosage for use in human. The dosage ofsuch compounds lies preferably within a range of circulatingconcentrations that include the ED₅₀ with little or no toxicity. Thedosage may vary within this range depending upon the dosage formemployed and the route of administration utilized. It is preferable thatthe therapeutic serum concentration of an efflux pump inhibitor shouldbe in the range of 0.1-100 μg/ml., more preferably 0.1-50 μg/ml.; 0.1-20μg/ml.; 1.0-50 μg/ml.; or 1.0-20 μg/ml.

For any compound used in the method of the invention, thetherapeutically effective dose can be estimated initially from cellculture assays. For example, a dose can be formulated in animal modelsto achieve a circulating plasma concentration range that includes theIC₅₀ as determined in cell culture Such information can be used to moreaccurately determine useful doses in humans. Levels in plasma may bemeasured, for example, by HPLC.

In particular preferred embodiments, the efflux inhibitor in apharmaceutical composition has a structure as shown by the genericstructures described above.

The exact formulation, route of administration and dosage can be chosenby the individual physician in view of the patient's condition. (Seee.g. Fingl et al., in THE PHARMACOLOGICAL BASIS OF THERAPEUTICS, 1975,Ch. 1 p. 1). It should be noted that the attending physician would knowhow to and when to terminate, interrupt, or adjust administration due totoxicity, or to organ dysfunctions. Conversely, the attending physicianwould also know to adjust treatment to higher levels if the clinicalresponse were not adequate (precluding toxicity). The severity of thecondition may, for example, be evaluated, in part, by standardprognostic evaluation methods. Further, the dose and perhaps dosefrequency, will also vary according to the age, body weight, andresponse of the individual patient. A program comparable to thatdiscussed above may be used in veterinary medicine.

Depending on the specific infection being treated, such agents may beformulated and administered systemically or locally. Techniques forformulation and administration may be found in Remington'sPharmaceutical Sciences, 18th ed., Mack Publishing Co., Easton, Pa.(1990). Suitable routes may include oral, rectal, transdermal, vaginal,transmucosal, or intestinal administration; parenteral delivery,including intramuscular, subcutaneous, intramedullary injections, aswell as intrathecal, direct intraventricular, intravenous,intraperitoneal, intranasal, or intraocular injections, just to name afew.

For injection, the agents of the invention may be formulated in aqueoussolutions, preferably in physiologically compatible buffers such asHanks's solution, Ringer's solution, or physiological saline buffer. Forsuch transmucosal administration, penetrants appropriate to the barrierto be permeated are used in the formulation. Such penetrants aregenerally known in the art.

Use of pharmaceutically acceptable carriers to formulate the compoundsherein disclosed for the practice of the invention into dosages suitablefor systemic administration is within the scope of the invention. Withproper choice of carrier and suitable manufacturing practice, thecompositions of the present invention, in particular, those formulatedas solutions, may be administered parenterally, such as by intravenousinjection. The compounds can be formulated readily usingpharmaceutically acceptable carriers well known in the art, into dosagessuitable for oral administration. Such carriers enable the compounds ofthe invention to be formulated as tablets, pills, capsules, liquids,gels, syrups, slurries, suspensions and the like, for oral ingestion bya patient to be treated.

Pharmaceutical compositions suitable for use in the present inventioninclude compositions wherein the active ingredients are contained in aneffective amount to achieve its intended purpose. Determination of theeffective amounts is well within the capability of those skilled in theart, especially in light of the detailed disclosure provided herein. Inaddition to the active ingredients, these pharmaceutical compositionsmay contain suitable pharmaceutically acceptable carriers includingexcipients and auxiliaries which facilitate processing of the activecompounds into preparations which can be used pharmaceutically. Thepreparations formulated for oral administration may be in the form oftablets, dragees, capsules, or solutions. The pharmaceuticalcompositions of the present invention may be manufactured in a mannerthat is itself known, e.g., by means of conventional mixing, dissolving,granulating, dragee-making, levitating, emulsifying, encapsulating,entrapping or lyophilizing processes.

Pharmaceutical formulations for parenteral administration includeaqueous solutions of the active compounds in water-soluble form.Additionally, suspensions of the active compounds may be prepared asappropriate oily injection suspensions. Suitable lipophilic solvents orvehicles include fatty oils such as sesame oil, or synthetic fatty acidesters, such as ethyl oleate or triglycerides, or liposomes. Aqueousinjection suspensions may contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.

Pharmaceutical preparations for oral use can be obtained by combiningthe active compounds with solid excipient, optionally grinding aresulting mixture, and processing the mixture of granules, after addingsuitable auxiliaries, if desired, to obtain tablets or dragee cores.Suitable excipients are, in particular, fillers such as sugars,including lactose, sucrose, mannitol, or sorbitol; cellulosepreparations such as, for example, maize starch, wheat starch, ricestarch, potato starch, gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/orpolyvinylpyrrolidone (PVP). If desired, disintegrating agents may beadded, such as the cross-linked polyvinyl pyrrolidone, agar, or alginicacid or a salt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers may be added.

EXAMPLES

The compounds of the present invention may be readily prepared inaccordance with the following synthesis schemes, as illustrated in thespecific examples provided. However, those skilled in the art willrecognize that other synthetic pathways for forming the compounds ofthis invention can be utilized, and that the following is providedmerely by way of example, and is not limiting to the present invention.It will be further recognized that various protecting and deprotectingstrategies will be employed which are standard in the art (see, e.g.,"Protective Groups in Organic Synthesis" by Greene and Wuts). Thoseskilled in the arts will recognize that the selection of any particularprotecting group (e.g., amine and carboxyl protecting groups) willdepend on the stability of the protected moiety with regard to thesubsequent reaction conditions and will understand the appropriateselections.

Further illustrating the knowledge of those skilled in the art is thefollowing sampling of the extensive chemical literature:

1) "Chemistry of the Amino Acids" by J. P. Greenstein and M. Winitz,Wiley and Sons, Inc. New York, N.Y. (1961).

2) "Comprehensive Organic Transformations" by R. Larock, VCH Publishers(1989)

3) T. D. Ocain and D. H. Rich, J. Med. Chem., 31, pp. 2193-2199 (1988).

4) E. M. Gordon, J. D. Godfrey, N. G. Delaney, M. M. Asaad, D. VonLangen, and D. W. Cushman, J. Med. Chem., 31, pp. 2199-2210 (1988).

5) "Practice of Peptide Synthesis" by M. Bodanszky and A. Bodanszky,Springer-Verlag, New York, N.Y. (1984).

6) "Protective Groups in Organic Synthesis" by T. Greene and P. Wuts(1991).

7) "Asymmetric Synthesis: Construction of Chiral Molecules Using AminoAcids" by G. M. Coppola and H. F. Schuster, John Wiley and Sons, NewYork, N.Y. (1987).

8) "The Chemical Synthesis of Peptides" J. Jones, Oxford UniversityPress, New York, N.Y. (1991).

9) "Introduction to Peptide Chemistry" by P. D. Bailey, John Wiley andSons, New York, N.Y. (1992).

10) "Synthesis of Optically Active α-Amino Acids" by R. M. Williams,Pergamon Press, Oxford, U.K. (1989). ##STR6## General Procedure forPhosphorus Oxychloride-Mediated Peptide Coupling Amidation (Procedure A)

A solution of N-protected amino acid in dichloromethane (0.1 M) at 0°C., under nitrogen atmosphere, is treated with phosphorus oxychloride(1.5 eq) and diisopropylethylamine (2.1 eq) followed by an alkyl (oraryl) amine (1.5 eq). The solution is stirred at 0° C. until startingmaterial was consumed, as per thin layer chromatography monitoring. Thereaction mixture is poured into ethyl acetate and worked up as usual,with purification by either chromatography or crystallization.

General Procedure for PyBrop-Mediated Peptide Coupling (Procedure B)

A solution of Nα-(alkylamino) component, Boc-amino acid (1.3 eq),diisopropylethyl-amine (2.0 eq), and dim ethylacetamide (6 ml) wastreated with benzotriazole-1-yl-oxytris(pyrrolidino)phosphoniumhexafluorophosphate (PyBrop) (1 eq) under nitrogen at room temperature.Reaction mixture is stirred 10-12 hrs, pour into ethyl acetate, andworked up as usual, with purification by either chromatography orcrystallization.

General Procedure for EDAC Mediated Peptide Coupling (Procedure C)

A solution of Boc-amino acid in dichloromethane (0.1 M),N-hydroxybenzotriazole (1 eq), and alkyl (or aryl) amine (1.6 eq) istreated with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (1.5eq).After stirring at room temperature for 10-12 hrs, the reaction mixtureis poured into ethyl acetate and work up as usual, with purification bychromatography or crystallization.

General Procedure for Coupling of N-Alkyl (or Arylalkyl) Peptides withMixed Anhydrides (Procedure D)

A cold (0° C.) solution of Boc-amino acid (1 mmol), triethylamine (1.2mmol), and dichloromethane (4 ml) under nitrogen atmosphere was treatedwith ethyl chloroformate [or pivaloyl chloride] (1.2 mmol). Afterstirring for 2 hrs at 0° C., a solution of secondary amine (1 mmol) indichloromethane (3.5 ml) was added and then the reaction mixture wasstirred at ambient temperature for 12-16 hrs. The reaction was worked upas in Procedure A.

General Procedure for Deprotection of tert-Butyloxycarbonyl (Boc)Peptides (Procedure E)

The starting material (10 mg) is dissolved in trifluoroacetic acid (1ml) and stirred 1 hr, and then concentrated in vacuo. The crude materialis loaded onto a reverse phase preparative HPLC. Typical HPLCconditions: 1 cm×22 cm Amberchrom; 2 ml/min flow. Solvent condition forone hour elution profiles: A: 0 to 50% acetonitrile/(0.1% TFA), B: 0 to60% acetonitrile/(0.1% TFA); C: 0 to 70% acetonitrile/(0.1% TFA). Thefractions are concentrated to remove acetonitrile, then lyophilized.

General Procedure for Reduction of N-Protected Amino Acids toN-Protected Amino Alcohols (Procedure F)

A cold solution (0° C.) of Boc-amino acid (1 mmol) in anhydroustetrahydrofuran (0.1 M), under nitrogen atmosphere, is treatedsequentially with ethyl chloroformate (2 eq) and triethylamine (2 eq).The mixture is stirred at 0° C. for 2 hours. Sodium borohydride (6 eq)is added, followed by very slow addition of water (16 ml) over a periodof 40 min. Once the addition is completed, the mixture is poured intoethyl acetate and worked up with final chromatographic purification.

Example 1 Alanyl-Phenylalanyl-Arginine 2-Naphthylamide Trifluoroacetate

A solution of Phe-Arg β-naphthylamide dihydrochloride (25 mg),diisopropylethylamine (8 μl), Boc-alanine N-hydroxysuccinimide ester (14mg), and dimethylacetamide (0.5 ml) was stirred at 25° C. for 2 hrs.After concentration in vacuo, resultant Boc-Ala-Phe-Arg 2-naphthylamidewas deprotected as described in Procedure E. Product was obtained aswhite solid (20 mg), after HPLC (method A, retention time=43.3 min.): ¹H NMR (400 MHz, D₂ O) δ 1.45 (d, J=8.6 Hz, 3H), 1.72 (m, 2H), 1.89 (m,1H), 2.00 (m, 1H), 3.15 (dd, J=12.9; 8.2 Hz, 1H), 3.24 (dd, J=12.3; 8.2Hz, 1H), 3.31 (t, J=7.1 Hz, 2H), 4.15 (q, J=8.8 Hz, 1H), 5.48 (t, J=8.0Hz, 1H), 4.81 (HOD with proton hidden), 7.20 (m, 1H), 7.29 (m, 4H), 7.58(d, J=10.6 Hz, 1H), 7.61 (m, 2H), and 7.99 (m, 4H).

Example 2 D-Alanyl-Phenylalanyl-Arginine 2-NaphthylamideTrifluoroacetate

This was similarly prepared, as described in Example 1. Boc-D-alanineN-hydroxy-succinimide ester was coupled to Phe-Arg-β-naphthylamidedihydrochloride; the resultant Boc-L-Ala-Phe-Arg 2-naphthylamide wasdeprotected with trifluoroacetic acid to afford a white solid: ¹ H NMR(400 MHz, D₂ O) δ 1.43 (d, J=8.8 Hz, 3H), 1.74 (m, 2H), 1.90 (m, 1H),1.99 (m, 1H), 3.17 (dd, J=12.9; 8.2 Hz, 1H), 3.21 (dd, J=12.3; 8.2 Hz,1H), 3.29 (t, J=7.1 Hz, 2H), 4.13 (q, J=8.8 Hz, 1H), 5.50 (t, J=8.0 Hz,1H), 4.81 (HOD with proton hidden), 7.18 (m, 1H), 7.29 (m, 4H), 7.58 (d,J=10.6 Hz, 1H), 7.63 (m, 2H), and 8.01 (m, 4H).

Example 3 D-Leucyl-Phenylalanyl-Arginine 2-NaphthylamideTrifluoroacetate

Using the procedure similar to that used in Example 1, Boc-D-leucineN-hydroxy-succinimide ester was coupled to Phe-Arg-β-naphthylamidedihydrochloride; the resultant Boc-L-Leu-Phe-Arg 2-naphthylamide wasdeprotected with trifluoroacetic acid to afford a white solid: ¹ H NMR(400 MHz, D₂ O) δ 0.85 (broad s, 6H), 1.29 (m, 1H), 1.52 (m, 2H), 1.77(m, 2H), 1.98 (m, 2H), 3.07 (m, 1H), 3.29 (m, 3H), 3.99 (m, 1H), 4.54(m, 1H), 4.81 (HOD with hidden proton), 7.22 (m, 1H), 7.36 (broad s,4H), 7.58 (d, J=10.0 Hz, 1H), 7.63 (m, 2H), 8.01 (m, 23H), and 8.09 (s,1H).

Example 4 Phenylalanyl-Ornithine Quinoline-3-amide Trifluoroacetate

(A) N-Boc-phenylalanyl-Nγ-Boc-ornithine

N-Boc-phenylalanine N-hydroxysuccinimide ester (1.3 g, 3.6 mmol) wasdissolved in dimethylformamide (15 mL) and Nγ-Boc-ornithine (0.88 g, 3.8mmol) was added in one portion. The solution was kept at 70° C. for 1hr, cooled to 25° C., filtered to clarify and concentrated in vacuo. Theresidue was dissolved in ethyl acetate and washed with water. Theorganic phase was dried over anhydrous sodium sulfate and concentratedto dryness to afford titled compound (1.02 g) as a white foam: ¹ H NMR(400 MHz, CDCl₃) δ 1.39-1.45 (18H), 1.70-1.72 (1H), 1.89-1.92 (1H),3.01-3.18 (4H), 4.43 (1H), 4.58 (1H), 4.82 (1H), 5.23 (1H), 7.20-7.32(5H).

(B) N-Boc-phenylalanyl-Nγ-Boc-ornithine Quinoline-3-amide

A cold solution (0° C.) of N-Boc-phenylalanyl-Nγ-Boc-ornithine (0.2 g,0.4 mmol), 3-amino-quinoline (0.082 g, 0.57 mmol), diisopropylethylamine(0.103 mg, 0.8 mmol), 4-(dimethyl-amino)pyridine (5 mg, 0.04 mmol), andmethylene chloride (3 mL) was treated dropwise with phosphorusoxychloride (0.5 mmol). The reaction was stirred at 0° C. for 1 hr andethyl acetate (20 mL) was added. The organic layer was washed with water(2×20 mL), 1N hydrochloric acid (2×10 mL), saturated sodium bicarbonate(2×10 mL) and brine. The organic layer was dried over anhydrous sodiumsulfate, filtered and the filtrate adsorbed onto 100 mg of silica geland applied to a column prepacked with silica gel. The column was elutedwith ethyl acetate/hexane (70:30, v:v) to afford titled compound (51 mg)as an oil: ¹ H NMR (400 MHz, CDCl₃) δ 1.42-1.55 (18H), 1.6-1.8 (1H),2.0-2.1 (1H), 3.1-3.2 (4H), 3.3-3.4 (1H), 4.19-4.21 (1H), 4.70-4.80(1H), 5.0-5.15 (1H), 7.21-7.29 (5H), 7.53-7.55 (1H), 7.63-7.65 (1H),7.79-7.81 (1H), 8.05-8.07 (1H), 8.74 (1H), 8.94 (1H); mass spectrum(relative intensity) m/e 606 (100, M+1).

(C) Phenylalanyl-Ornithine Quinoline-3-amide Trifluoroacetate

A solution of N-Boc-phenylalanyl-Nγ-Boc-ornithine quinoline-3-amide (50mg) and trifluoroacetic acid (2.5 mL) was stirred at 25° C. for 1 hr.The solution was concentrated in vacuo, suspended in water and appliedto a MPLC reverse phase column (1 cm×22 cm, Amberchrom). The column waseluted at a rate of 2 mL/min over 1 hr (gradient of 0 to 60%acetonitrile with 0.1% TFA) and desired fractions lyophilized to affordtitled dipeptide amide (46 mg): ¹ H NMR (400 MHz, D₂ O) δ 1.83-2.15(4H), 3.11-3.15 (2H), 3.27-3.34 (2H), 4.39-4.43 (1H), 4.63-4.67 (1H),7.17-7.34 (5H), 8.01-8.05 (1H), 8.12-8.16 (2H), 9.03 (1H), 9.37 (1H);mass spectrum (relative intensity) m/e 406 (100, M+1).

Example 5 β-N-(Phenethyl)alanine Methyl Ester

A mixture of methyl acrylate (2.0 g), phenethylamine (3.1 g), anhydrousmethanol (100 ml), and glacial acetic acid (100 mg) was stirred at 25°C. for 14 hr, concentrated in vacuo and the resultant oil adsorbed ontosilica gel (5 g) and applied to a column prepacked with silica gel. Thetitle compound (2.2 g) was eluted from the column with CH₂ Cl₂ :MeOH:NH₄OH (89:9:2, v:v): ¹ H NMR (400 MHz, CDCl₃) δ 2.50-2.53 (2H), 2.79-2.94(6H), 3.66 (3H), 7.20-7.32 (5H).

Example 6 β-N-(3-Phenylpropyl)alanine Methyl Ester

This was similarly prepared, as described in Example 5, except thestarting materials are methyl acrylate and 3-phenylpropylamine.

Example 7 β-N-(p-Tolylethyl)alanine Ethyl Ester

This was similarly prepared, as described in Example 5, except thestarting materials are ethyl acrylate and p-tolylethylamine.

Example 8 β-N-(iso-Butyl)alanine Methyl Ester

This was similarly prepared, as described in Example 5, except thestarting materials are methyl acrylate and iso-butylamine.

Example 9 β-N-(Cyclohexylmethyl)alanine Ethyl Ester

This was similarly prepared, as described in Example 5, except thestarting materials are ethyl acrylate and cyclohexylmethylamine.

Example 10 β-N-(4-Fluorophenylpropyl)alanine Methyl Ester

This was similarly prepared, as described in Example 5, except thestarting materials are methyl acrylate and 4-fluorophenylpropylamine.

Example 11 β-N-(Cyclopropylmethyl)alanine Methyl Ester

This was similarly prepared, as described in Example 5, except thestarting materials are methyl acrylate and cyclopropylmethylamine.

Example 12 β-N-(3-Ethoxypropyl)alanine Methyl Ester

This was similarly prepared, as described in Example 5, except thestarting materials are methyl acrylate and 3-ethoxypropylamine.

Example 13 D-Ornithyl-β-N-(3-Phenylpropyl)alanine Quinoline-3-amideTrifluoroacetate

This was prepared, as in Example 4, in two steps. Initial coupling ofNα,Nγ-bis-Boc-D-ornithine and methyl β-N-(3-phenylpropyl)alaninate(Procedure B) affordedNα,Nγ-Bis-Boc-D-ornithyl-β-(3-phenylpropyl)alanine methyl ester.Subsequent hydrolysis (0.1N sodium hydroxide), coupling with3-aminoquinoline (Procedure A), and deprotection (Procedure E) gave thetitled compound.

Example 14 D-Ornithyl-β-N-(3-Phenylpropyl)alanine 2-NaphthylamideTrifluoroacetate

This was prepared, as in Example 13, except the starting materials wereNα,Nγ-bis-Boc-D-ornithine, methyl β-N-(3-phenylpropyl)alaninate, and2-aminonaphthalene.

Example 15 D-Ornithyl-β-N-(iso-Butyl)alanine Quinoline-3-amideTrifluoroacetate

This was prepared, as in Example 13, except the starting materials wereNα,Nγ-bis-Boc-D-ornithine, methyl β-N-(iso-butyl)alaninate, and3-aminoquinoline.

Example 16 D-Lysyl-β-N-(iso-Butyl)alanine Quinoline-3-amideTrifluoroacetate

This was prepared, as in Example 13, except the starting materials wereNα,Nε-bis-Boc-D-lysine, methyl β-N-(iso-butyl)alaninate, and3-aminoquinoline.

Example 17 D-Lysyl-β-N-(iso-Butyl)alanine (3-Phenylpropyl)amideTrifluoroacetate

This was prepared, as in Example 13, except the starting materials wereNα,Nε-bis-Boc-D-ysine, methyl β-N-(iso-butyl)alaninate, and3-phenylpropylamine.

Example 18 D-Lysyl-β-N-(Cyclohexylmethyl)alanine Quinoline-3-amideTrifluoroacetate

This was prepared, as in Example 13, except the starting materials wereNα,Nε-bis-Boc-D-lysine, ethyl β-N-(cyclohexylmethyl)alaninate, and3-aminoquinoline.

Example 19 D-Ornithyl-β-N-(Cyclohexylmethyl)alanine 2-NaphthylamideTrifluoroacetate

This was prepared, as in Example 13, except the starting materials wereNα,Nγ-bis-Boc-D-ornithine, ethyl β-N-(cyclohexylmethyl)alaninate, and2-aminonaphthalene.

Example 20 D-Arginyl-β-N-(iso-Butyl)alanine Quinoline-2-amideTrifluoroacetate

This was prepared, as in Example 13, except the starting materials wereNα,Nγ,Nγ-tri-Boc-D-arginine, methyl β-N-(iso-butyl)alaninate, and2-aminoquinoline.

Example 21 D-Lysyl-β-N-(iso-Butyl)alanine Quinoline-3-amideTrifluoroacetate

This was prepared, as in Example 13, except starting materials wereNα,Nε-bis-Boc-D-lysine, methyl β-N-(iso-butyl)alaninate, and3-aminoquinoline.

Example 22 D-Lysyl-β-N-(4-Methylphenethyl)alanine Quinoline-2-amide TFA

This was prepared, as in Example 13, except the starting materials wereNα,Nε-bis-Boc-D-lysine, methyl β-N-(4-methylphenethyl)alaninate, and2-aminoquinoline.

Example 23 D-Ornithyl-β-N-(4-Methylphenethyl)alanine Quinoline-3-amideTFA

This was prepared, as in Example 13, except starting materials wereNα,Nγ-bis-Boc-D-ornithine, methyl β-N-(4-methylphenethyl)alaninate, and3-aminoquinoline.

Example 24 D-Ornithyl-β-N-(Ethylthioethyl)alanine 2-NaphthylamideTrifluoroacetate

This was prepared, as in Example 13, except the starting materials wereNα,Nγ-bis-Boc-D-ornithine, methyl β-N-(ethylthioethyl)alaninate, and2-aminonaphthalene.

Example 25 D-Lysyl-β-N-(Ethylthioethyl)alanine 2-NaphthylamideTrifluoroacetate

This was prepared, as in Example 13, except starting materials wereNα,Nε-bis-Boc-D-lysine, methyl β-N-(ethylthioethyl)alaninate, and2-aminonaphalene.

Example 26 D-Lysyl-β-N-(Ethylthioethyl)alanine 2-NaphthylamideTrifluoroacetate

This was prepared, as in Example 13, except starting materials wereNα,Nε-bis-Boc-D-lysine, methyl β-N-(ethylthioethyl)alaninate, and3-aminoquinoline.

Example 27 D-Lysyl-β-N-(Cyclopropylmethyl)alanine Quinoline-3-amideTrifluoroacetate

This was prepared, as in Example 13, except starting materials wereNα,Nε-bis-Boc-D-lysine, methyl β-N-(cyclopropylmethyl)alaninate, and3-aminoquinoline.

Example 28 D-Ornithyl-β-N-(Cyclopropylmethyl)alanine Quinoline-2-amideTrifluoroacetate

This was prepared, as in Example 13, except the starting materials wereNα,Nγ-bis-Boc-D-ornithine, methyl β-N-(cyclopropylmethyl)alaninate, and2-aminoquinoline.

Example 29 D-Lysyl-β-N-(Cyclopropylmethyl)alanine Quinoline-2-amideTrifluoroacetate

This was prepared, as in Example 13, except the starting materials wereNα,Nε-bis-Boc-D-lysine, methyl β-N-(cyclopropylmethyl)alaninate, and2-aminoquinoline.

Example 30 D-Lysyl-β-N-(3,3-Dimethylbutyl)alanine 2-NaphthylamideTrifluoroacetate

This was prepared, as in Example 13, except starting materials wereNα,Nε-bis-Boc-D-lysine, methyl β-N-(3,3-dimethylbutyl)alaninate, and2-aminonaphthalene.

Example 31 Ornithyl-β-N-(3-Phenylpropyl)alanine 2-NaphthylamideTrifluoroacetate

This was prepared, as in Example 13, except starting materials wereNα,Nγ-bis-Boc-ornithine, methyl β-N-(3-phenylpropyl)alaninate, and2-aminonaphthalene.

Example 32 Lysyl-β-N-(3-Phenylpropyl)alanine Quinoline-3-amideTrifluoroacetate

This was prepared, as in Example 13, except starting materials wereNα,Nε-bis-Boc-lysine, methyl β-N-(3,3-dimethylbutyl)alaninate, and3-aminoquinoline.

Example 33 D-Ornithyl-D-Phenylalanine Quinoline-3-amide Trifluoroacetate

(A) N-Boc-D-phenylalanine Quinoline-3-amide

A solution of N-Boc-D-phenylalanine (1.25 g, 4.7 mmol) in ethyl acetate(40 mL) was treated sequentially with 3-aminoquinoline (1.4 g, 9.4 mmol)and dicyclohexylcarbo-diimide (1.02 g, 4.9 mmol). The reaction mixturewas stirred at room temperature for 16 hr, filtered and the filtratewashed with 1M hydrochloric acid (2×25 mL), saturated sodium bicarbonate(1×25 mL), and brine (1×25 mL). The organic layer was dried overanhydrous sodium sulfate, filtered and concentrated to dryness to affordtitled compound (950 mg) as an oil.

(B) Nα,Nγ-Boc-D-ornithyl-D-Phenylalanine Quinoline-3-amide

A solution of Nα,Nγ-Boc-D-ornithine (253 mg, 0.76 mmol), triethylamine(81 mg, 0.8 mmol), and methylene chloride (10 ml) was stirred at 25° C.for 10 min, cooled to 0° C. and treated with ethyl chloroformate (82 mg,0.76 mmol) of ethyl chloroformate added. The mixture was stirred at 0°C. for 2.5 hr. During this time, N-Boc-phenylalanine quinoline-3-amide(200 mg, 0.51 mmol) was treated with trifluoroacetic acid (5 mL) at 25°C. for 45 min. The solution was concentrated to dryness, coevaporatedwith methylene chloride (3×5 mL), redissolved in methylene chloride (10mL) and neutralized to pH8 with triethyl-amine (2 eq.). This solutionwas added to the mixed anhydride and the mixture was stirred at roomtemperature for 2 hr at which time the reaction was quenched by theaddition of sat. sodium bicarbonate (20 mL). The organic layer waswashed with brine (10 mL), dried over anhydrous sodium sulfate, andconcentrated to dryness to afford Nα,Nγ-Boc-D-ornithyl-D-phenylalaninequinoline-3-amide as a white solid.

(C) D-Ornithyl-D-Phenylalanine Quinoline-3-amide Trifluoroacetate

Nα,Nγ-Boc-D-ornithyl-D-phenylalanine quinoline-3-amide was treated withtrifluoro-acetic acid (20 mL) at 25° C.; after 1 hr, the reaction wasconcentrated in vacuo and the residue was purified by reverse-phasechromatography (Amberchrom) to afford titled compound as white solid:

Example 34 D-Ornithyl-D-β-(3-Quinolinyl)alanine (3-Phenylpropyl)amide

This was similarly prepared, as described in Example 33, except thestarting materials are Boc-D-β-(3-quinolinyl)alanine,Nα,Nγ-bis-Boc-D-ornithine, and 3-phenylpropylamine.

Example 35 D-Ornithyl-D-β-(3-Quinolinyl)alanine (4-ethylbenzyl)amide

This was similarly prepared, as described in Example 33, except thestarting materials are Boc-D-β-(3-quinolinyl)alanine,Nα,Nγ-bis-Boc-D-ornithine, and 4-ethylbenzylamine.

Example 36 D-Ornithyl-D-β-(3-Quinolinyl)alanine2,3-trimethylenepyridyl-5-amide

This was similarly prepared, as described in Example 33, except thestarting materials are Boc-D-β-(3-quinolinyl)alanine,Nα,Nγ-bis-Boc-D-ornithine, and 5-amino-2,3-trimethy-lenepyridine.

Example 37 D-Lysyl-D-β-(3-Quinolinyl)alanine Isobutylamide

This was similarly prepared using the procedure in Example 33, exceptthe starting materials are Boc-D-β-(3-quinolinyl)alanine,Nα,Nγ-bis-Boc-D-lysine, and i-butylamine.

Example 38 Phenylalanyl-Nα-Methylarginine β-NaphthylamideTrifluoroacetate

(A) Nα-Boc-Nγ-Fmoc-Nα-Methylornithine

Compound is prepared using a revised literature procedure (C.-B. Xue andW. F. DeGrado, Tetrahedron Lett., 36, 55 (1995), but using Fmoc-Clinstead of Cbz-Cl: ¹ H NMR (400 MHz, CDCl₃) δ 1.45 (s, 9 H), 1.60-2.15(m, 4 H), 2.80 (s, 3I), 3.00 (m, 2 H), 4.50 (m, 2 H), 4.18 (m, 1 H),4.21 (m, 1 H), 4.42 (m, 2 H), 7.33 (m, 2 H), 7.39 (m, 2 H), 7.59 (m, 2H), and 7.78 (d, J=8.9 Hz, 2 H).

(B) Nα-Boc-Nγ-Fmoc-Nα-Methylornithine β-Naphthylamide

This compound is prepared using Procedure A.Nα-Boc-Nγ-Fmoc-Nα-methylomithine (80 mg), obtained in (A), andβ-naphthylamine were condensed to afford a colorless solid (103 mg): ¹ HNMR (400 MHz, CDCl₃) δ 1.53 (s, 9 H), 1.85 (m, 2 H), 2.15 (m, 1 H), 2.94(s, 3 H), 3.30 (m, 2 H), 4.25 (t, J=6.4 Hz, 1 H), 4.42 (d, 2 H), 4.82(s, 1 H), 5.00 (s, 1 H), 7.30 (m, 2 H), 7.44 (m, 8 H), 7.76 (m, 2 H),7.78 (m, 2 H), and 8.25 (s, 1 H).

(C) Boc-Phenylalanyl-Nγ-Fmoc-Nα-Methylornithine β-Naphthylamide

This compound is prepared using Procedure D from Boc-phenylalanine (49mg) and Nγ-Fmoc-Nα-methylornithine-2-naphthylamide (100 mg) to give aglassy solid (125 mg): ¹ H NMR (400 MHz, CDCl₃) δ 1.39 (s, 9 H), 1.51(m, 2 H), 1.78 (m, 1 H), 1.89 (m, 1 H), 2.85 (s, 3 H), 2.95-3.18 (m, 2H), 4.25 (m, 1 H), 4.40 (m, 2 H), 4.83 (m, 1 H), 5.05 (m, 1 H), 7.25 (m,5 H), 7.30 (m, 4 H), 7.32 (m, 4 H), 7.43 (m, 4 H), 7.60 (m, 1 H), and7.80 (m, 2 H).

(D) Boc-Phenylalanyl-Nω,Nω'-bis-Boc-Nα-Methylarginine β-Naphthylamide

This compound is prepared in two steps by first dissolvingBoc-phenylalanyl-Nγ-Fmoc-Nα-methylornithine β-naphthylamide (100 mg),obtained from (C), in 20% piperidine in dimethyl-acetamide (5 ml),stirring 20 min. at ambient temperature, concentrating and drying undervacuum. The residue is then dissolved in dimethylformamide (5 ml)followed by the addition of N,N'-bis-Boc-1-guanylpyrazole (Y. Wu, G. R.Matsueda, M. Bernatowicz, Synth. Comm., 23, 3055 (1993); 42 mg) anddiisopropylethylamine (71 μl). The reaction mixture is poured into ethylacetate, worked up as usual and the desired compound purified by flashchromatography to give titled product (103 mg) as a white solid. Thecompound appears as a 1:1-mixture of rotamers: ¹ H NMR (400 MHz, CDCl₃)δ 1.41-1.90 (m, 31 H), 2.67, 2.80 (2s, total 3 H), 2.91-3.10 (m, 2 H),3.45 (m, 2 H), 4,22 (m), 4.61 (dd, J=11.4; 3.6 Hz, 1 H), 4.86 (m, 2 H),7.20 (m, 3 H), 7.25 (m, 5 H), 7.43 (m, 2 H), and 7.83 (m, 2 H).

(E) Phenylalanyl-Nα-Methylarginine β-Naphthylamide Trifluoroacetate

Boc-Phenylalanyl-Nω,Nω'-bis-Boc-Nα-methylarginine β-naphthylamide wastreated with trifluoroacetic acid (Procedure E), followed by HPLCpurification (Method A) to afford a white solid (45 mg): ¹ H NMR (400MHz, D₂ O) δ 1.70 (m, 2 H), 1.92 (m, 1 H), 2.15 (m, 1 H), 2.82 (s, 3H),3.42 (m, 4 H), 4.83 (HOD with proton hidden), 5.19 (m, 1 H), 7.18 (m, 2H), 7.31 (m, 1 H), 7.68 (m, 4 H), 8.15 (m, 4 H), and 8.20 (s, 1 H).

Example 39 Phenylalanyl-Nα-Methylornithine β-NaphthylamideTrifluoroacetate

A solution of Boc-phenylalanyl-Nγ-Fmoc-Nα-methylornithineβ-naphthylamide (60 mg), in 20% piperidine in dimethylacetamide (5 ml),was stirred for 20 min. at 25° C., concentrated and dried under vacuum.Crude Boc-phenylalanyl-Nα-methylornithine β-naphthylamide is thendeprotected, as per Procedure E, to give a white solid which waspurified by HPLC (method A): ¹ H NMR (400 MHz, D₂ O) δ 1.74 (m, 2 H),1.87 (m, 1 H), 2.10 (m, 1 H), 2.78 (s, 3 H), 3.11 (t, J=7.6 Hz, 2 H),3.21 (dd, J=13.2, 8.8 Hz, 1 H), 3.38 (dd, J=13.2; 5.2 Hz, 1 H), 4.89(HOD with proton hidden), 5.22 (t, J=7.6 Hz, 1 H), 7.15 (m, 3 H), 7.28(m, 2 H), 7.68 (m, 3 H), 8.04 (m, 3 H), and 8.13 (s, 1 H).

Example 40 Phenylalanyl-N.sub.α -Methylornithine (2-Naphthyl)methylamideTrifluoroacetate

(A) Nα-Boc-Nγ-Fmoc-Nα-Methylornithine (2-Naphthyl)methylamide

This compound is prepared using Procedure A; fromNα-Boc-Nγ-Fmoc-Nα-methylornithine and (2-naphthyl)methylamine to afforda colorless solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.42 (s, 9H), 1.56 (m, 2H), 1.78 (m, 1 H), 1.95 (m, 1 H), 2.82 (s, 3 H), 3.28 (m, 2 H), 4.21 (m,1 H), 4.39 (m, 2 H), 4.51 (m, 1 H), 4.69 (m, 3 H), 7.40 (m, 5 H), 7.51(m, 2 H), 7.58 (d, J=5.3 Hz, 2 H), 7.68 (s, 1 H), and 7.81 (m, 5 H).

(B) Boc-Phenylalanyl-Nγ-Fmoc-Nα-Methylornithine (2-Naphthyl)methylamide

This compound is prepared in two steps.Nα-Boc-Nγ-Fmoc-Nα-methylornithine (2-naphthyl)-methylamide (80 mg),obtained from (A), is deprotected with trifluoroacetic acid (5 ml),concentrated and coevaporated thrice with toluene. The crude residue isthen neutralized with triethylamine in dichloromethane and coupled toBoc-phenylalanine, using Procedure D, to give title compound (24 mg) asa glassy solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.50 (s, 9 H), 1.79 (m, 2 H),2.03 (m, 2 H), 2.82 (s, 3 H), 3.23 (m, 4 H), 4.18 (m, 3 H), 4.39 (m, 2H), 4.59 (dd, J=13.3; 6.1 Hz, 1 H), 4.64 (dd, J=13.0; 5.5 Hz, 1 H), 7.23(m, 8 H), 7.42 (m, 2 H), 7.57 (m, 2 H), 7.66 9 s, 1 H), and 7.80 (m, 7H).

(C) Phenylalanyl-Nα-Methylornithine (2-Naphthyl)methylamideTrifluoroacetate

A solution of Boc-phenylalanyl-Nγ-Fmoc-Nα-methylornithine(2-naphthyl)methylamide (24 mg) and 20% piperidine in dimethylacetamide(1.5 ml) was stirred for 20 min. at 25° C., and concentrated in vacuo.The residue is further deprotected, as per Procedure E, to give desiredproduct (14 mg) as a white solid, HPLC (method A): ¹ H NMR (400 MHz, D₂O) δ 1.72 (m, 2 H), 1.85 (m, 1 H), 2.10 (m, 1 H), 2.82 (s, 3 H), 3.12(m, 4 H), 4.57 (d, J=13.2 Hz, 1 H), 4.75 (d, J=13.1 Hz, 1 H), 4.80 (HODwith proton hidden), 5.09 (t, J=9.5 Hz, 1 H), 7.08 (m, 2 H), 7.24 (m, 3H), 7.59 (m, 4 H), and 7.98 (m, 3 H).

Example 41 Phenylalanyl-Nα-Methylornithine 2,2-DiphenylethylamideTrifluoroacetate

(A) Nα-Boc-Nγ-Fmoc-Nα-Methylornithine 2,2-Diphenylethylamide

Using Procedure A, Nα-Boc-Nγ-Fmoc-Nα-methylornithine and2,2-diphenylethylamine afforded a colorless solid: ¹ H NMR (400 MHz,CDCl₃) δ 1.39 (s, 10 H), 1.57 (m, 2 H), 1.77 (m, 1 H), 1.98 (m, 1 H),2.50 (s, 3 H), 3.19 (m, 2 H), 3.80 (m, 1 H), 4.00 (m, 1 H), 4.19 (t,J=9.5 Hz, 1 H), 4.22 (m, 1 H), 4.40 (d, J=7.2 Hz, 2 H), 4.46 (m, 1 H),7.20-7.34 (m, 12 H), 7.41 (t, J=6.6 Hz, 2 H), 7.60 (d, J=8.0 Hz, 2 H),and 7.79 (d, J=8.5 Hz, 2 H).

(B) Phenylalanyl-Nα-Methylornithine 2,2-Diphenylethyl amideTrifluoroacetate.

Nα-Boc-Nγ-Fmoc-Nα-methylornithine 2,2-diphenylethylamide (A) wasconverted in two steps (similar to that exemplified in Example 4) to awhite solid: ¹ H NMR (400 MHz, D₂ O) δ 1.51 (m, 2 H), 1.72 (m, 1 H),1.85 (m, 1 H), 2.72 (s, 3 H), 2.91 (m, 2 H), 3.12 (t, J=6.1 Hz, 2 H),3.94 (dd, J=12.8, 9.1 Hz, 1 H), 4.09 (dd, J=12.8, 9.0 Hz, 1 H), 4.41 (t,J=7.9 Hz, 1 H), 4.68 (t, J=9.6 Hz, 1 H), 4.95 (t, J=9.1 Hz, 1 H), and7.22-7.53 (m, 15 H).

Example 42 β-(4-Fluorophenyl)alanyl-Nα-Methylornithine β-NaphthylamideTrifluoroacetate

(A) Boc-β-(4-Fluorophenyl)alanyl-Nγ-Boc-Nα-Methylornithineβ-Naphthylamide

Using Procedure D; Boc-β-(4-fluorophenyl)alanine (138 mg) andNγ-Boc-Nα-methyl-ornithine β-naphthylamide (120 mg) afforded the titledcompound (184 mg) as a glassy solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.41 (s,18 H), 1.62 (m, 2 H), 1.80 (m, 1 H), 2.02 (m, 1 H), 2.82 (s, 3 H),2.92-3.20 (m, 4 H), 4.61 (m, 1 H), 4.82 (m, 1 H), 6.70 (m, 1 H), 7.05(m, 2 H), 7.22 (m, 1 H), 7.41 (m, 3 H), 7.79 (m, 3 H), and 8.20 (s, 1H).

(B) β-(4-Fluorophenyl)alanyl-Nα-Methylornithine β-NaphthylamideTrifluoroacetate

Deprotection of Boc-β-(4-fluorophenyl)alanyl-Nγ-Boc-Nα-methylornithineβ-naphthyl-amide (174 mg) with trifluoroacetic acid (Procedure E)afforded titled compound as a white solid (161 mg): HPLC (method A,retention time=42.27 min); ¹ H NMR (400 MHz, D₂ O) δ 1.76 (m, 2 H), 1.90(m, 1 H), 2.12 (m, 1 H), 2.84 (s, 3 H), 3.13 (t, J=7.6 Hz, 2 H), 3.21(dd, J=9.2; 13.6 Hz, 1 H), 3.37 (dd, J=4.8; 13.2 Hz, 1 H), 4.88 (t,J=9.6 Hz, 1 H), 5.23 (t, J=7.6 Hz, 1 H), 6.81 (t, J=8.4 Hz, 2 H), 7.29(m, 2 H), 7.63 (m, 3 H), 8.04 (m, 3 H), and 8.14 (s, 1 H).

Example 43 Tyrosyl-Nα-Methylornithine β-Naphthylamide Trifluoroacetate

(A) Boc-Tyrosyl-Nγ-Boc- Nα-Methylornithine β-Naphthylamide

Boc-tyrosine (400 mg) and Nγ-Boc-Nα-methylornithine β-naphthylamide (454mg) were coupled (Procedure B) to afford titled compound (447 mg) as aglassy solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.50 (m, 21 H), 1.77 (m, 1 H),2.73 (s, 3 H), 2.76 (m, 2 H), 2.99 (dd, J=10.8; 6.8 Hz, 1 H), 3.09 (dd,J=12.8; 13.2 Hz, 1 H), 4.70 (m, 1 H), 4.87 (m, 1 H), 6.89 (d, J=8.0 Hz,2 H), 7.09 (d, J=8.0 Hz, 2 H), 7.41 (m, 2 H), 7.75 (m, 4 H), and 8.26(s, 1 H).

(B) Tyrosyl-Nα-Methylornithine β-Naphthylamide Trifluoroacetate

Using Procedure E, Boc-tyrosyl-Nγ-Boc-Nα-methylomithine β-naphthylamide(A) (43 mg) afforded the desired compound as a white solid (36 mg): HPLC(method A, retention time=38.81 min); ¹ H NMR (400 MHz, D₂ O) δ 1.78 (m,2 H), 1.88 (m, 1 H), 2.10 (m, 1 H), 2.87 (s, 3 H), 3.13 (m, 3 H), 3.31(dd, J=14.0; 5.6 Hz, 1 H), 4.81 (HOD with proton hidden), 5.23 (t, J=7.6Hz, 1 H), 6.58 (d, J=8.4 Hz, 2 H), 7.18 (d, J=8.4 Hz, 2 H), 7.59 (dd,J=9.2; 2.0 Hz, 1 H), 7.66 (m, 2 H), 8.03 (m, 2 H), 8.08 (d, J=8.8 Hz, 1H), and 8.12 (d, J=1.6 Hz, 1 H); mass spectrum (ES+) m/e 435 (M+1).

Example 44 Homophenylalanyl-Nα-Methylornithine2-(4-Fluorophenyl)ethylamide Trifluoroacetate

(A) Nγ-Boc-Nα-Benzyl-Nα-Methylornithine 2-(4-Fluorophenyl)ethylamide

Nγ-Boc-Nα-benzyl-Nα-methylornithine and 2-(4-fluorophenyl)ethylamidewere coupled, as per Procedure C, to afford titled compound as a glassysolid: ¹ H NMR (400 MHz, CDCl₃) δ 1.52 (s, 9 H), 1.73 (m, 1 H), 1.65 (m,2 H), 1.79 (m, 1 H), 2.78 (t, J=7.9 Hz, 2 H), 3.02 (m, 1 H), 3.14 (m, 2H), 3.52 (m, 4 H), 6.96 (m, 2 H), 7.11 (m, 5 H), and 7.28 (m, 2 H).

(B) Nγ-Boc-Nα-Methylornithine 2-(4-Fluorophenyl)ethylamide

Hydrogen gas was bubbled through a solution ofNγ-Boc-Nα-benzyl-Nα-methylornithine 2-(4-fluorophenyl)ethylamide (A)(180 mg) in methanol (10 ml) in the presence 10% palladium-on-charcoal(20 mg). After starting material had disappeared, as per thin-layerchromatography monitoring, the reaction mixture was filtered through a0.45 μm nylon pad and concentrated. The product is used crude: ¹ H NMR(400 MHz, CDCl₃) δ 1.42 (m, 11 H), 1.50 (m, 2 H), 2.28 (s, 3 H), 2.80(t, J=7.5 Hz, 2 H), 2.94 (t, J=6.0 Hz, 1H), 3.11 (m, 2 H), 3.53 (q,J=8.1 Hz, 2 H), 6.98 (t, J=9.7 Hz, 2 H), and 7.18 (dd, J=9.7; 8.5 Hz, 2H).

(C) Boc-Homophenylalanyl-Nγ-Boc-Nα-Methylornithine2-(4-Fluorophenyl)ethylamide

Using Procedure D, Boc-homophenylalanine (143 mg) andNγ-Boc-Nα-methylornithine 2-(4-fluorophenyl)ethylamide (B) (crudeproduct) gave desired compound (195 mg) as a glassy solid which waspurified by flash chromatography: ¹ H NMR (400 MHz, CDCl₃) δ 1.52 (2 s,18 H) 1.71 (m, 2 H), 1.82 (m, 4 H), 2.62-2.80 (m, 7 H), 3.12 (m, 2 H),3.45 (m, 2 H), 4.61 (m, 1 H), 4.92 (m, 1 H), 6.89 (m, 2 H), 7.13 (m, 2H), 7.20 (m, 3 H), and 7.32 (m, 2 H).

(D) Homophenylalanyl-Nα-Methylornithine (4-Fluorophenyl)ethylamideTrifluoroacetate

Titled compound is obtained fromBoc-homophenylalanyl-Nγ-Boc-Nα-methylornithine2-(4-fluorophenyl)ethylamide (Procedure E): HPLC (method A); ¹ H NMR(400 MHz, D₂ O) δ 1.59 (m, 2 H), 1.78 (m, 1 H), 1.88 (m, 1 H), 2.15 (m,2),2.78 (s, 3 H), 2.83 (m, 2 H), 3.09 (m, 2 H), 3.58 (m, 2H), 4.48 (m, 1H), 4.95 (m, 1 H), 7.12 (m, 2 H), 7.23 (m, 2 H), 7.38 (m, 3 H), and 7.51(m, 2 H).

Example 45 β-(4-Iodophenyl)alanyl-Nα-Methylornithine(4-Fluorophenyl)ethylamide Trifluoroacetate

(A) Boc-β-(4-Iodophenyl)alanyl-Nγ-Boc-Nα-Methylornithine2-(4-Fluorophenyl)ethyl-amide

Boc-β-(4-iodophenyl)alanine and Nγ-Boc-Nα-methylornithine2-(4-fluorophenyl)ethyl-amide were coupled by Procedure D: ¹ H NMR (400MHz, CDCl₃) δ 1.38-1.51 (m, 20 H), 1.82 (m, 1 H), 1.90 (m, 1 H), 2.78(s, 3 H), 2.80-2.97 (m, 4 H), 3.00-3.17 (m, 2 H), 3.28-3.44 (m, 2 H),4.63 (m, 1 H), 4.97 (m, 1 H), 6.94-6.99 (m, 4 H), 7.11 (m, 1 H), 7.18(m, 1 H), 7.58 (d, 1 H), and 7.63 (dd, 1 H).

(B) β-(4-Iodophenyl)alanyl-Nα-Methylornithine2-(4-Fluorophenyl)ethylamide Trifluoroacetate

Boc-β-(4-iodophenyl)alanyl-Nγ-Boc-Nα-methylornithine2-(4-fluorophenyl)ethylamide was deprotected with trifluoroacetic acid(Procedure E) to afford a white solid: ¹ H NMR (400 MHz, D₂ O) δ 1.50(m, 1 H), 1.60 (m, 1 H), 1.71 (m, 1 H), 1.83 (m, 1 H), 2.73 (s, 2 H),2.92 (m, 2 H), 3.03 (m, 3 H), 3.12 (m, 1 H), 3.59 (m, 2 H), 4.74 (m, 1H), 4.91 (m, 1 H), 7.04 (d, J=8.0 Hz, 1 H), 7.11 (t, J=10.9 Hz, 2 H),7.35 (m, 2 H), and 7.79 (d, J=8.5 Hz, 2 H).

Example 46 Homophenylalanyl-Nα-Methylornithine2-(4-Methylphenyl)ethylamide Trifluoroacetate

(A) Nα-Benzyl-Nγ-Boc-Nα-Methylornithine 2-(4-Methylphenyl)ethylamide

Nα-Benzyl-Nγ-Boc-Nα-methylornithine (200 mg) and2-(4-methylphenyl)ethylamine were coupled to afford the titled compound(108 mg) as a glassy solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.45 (s, 9 H),1.58 (m, 1 H), 1.65 (m, 2 H), 1.75 (m, 1 H), 2.13 (s, 3 H), 2.35 (s, 3H), 2.81 (t, J=5.8 Hz, 2 H), 3.02 (m, 1 H), 3.15 (m, 2 H), 3.58 (m, 4H), 7.09 (m, 6 H), and 7.27 (m, 3 H).

(B) Nγ-Boc-Nα-Methylornithine 2-(4-Methylphenyl)ethylamide

Catalytic reduction of Nα-benzyl-Nγ-Boc-Nα-methylornithine2-(4-methylphenyl)ethyl-amide afforded the titled compound: ¹ H NMR (400MHz, CDCl₃) δ 1.43 (s, 9 H), 1.51 (m, 3 H), 1.64 (m, 1 H), 2.31 (s, 3H), 2.33 (s, 3 H), 2.80 (t, J=8.6 Hz, 1 H), 2.95 (m, 1 H), 3.11 (m, 2H), 3.52 (m, 2 H), 4.68 (m, 1 H), and 7.12 (m, 4 H).

(C) Boc-Homophenylalanyl-Nγ-Boc-Nα-Methylornithine2-(4-Methylphenyl)ethylamide

Using Procedure D, Boc-homophenylalanine (143 mg) andNγ-Boc-Nα-methylornithine 2-(4-methylphenyl)ethylamide (crude productfrom B) afforded titled product (195 mg) as a glassy solid after silicagel chromatography: ¹ H NMR (400 MHz, CDCl₃) δ 1.44 (2 s, 18 H), 1.63(m, 2 H), 1.83 (m, 2 H, 2.33 s, 3 H), 2.63-2.79 (m, 7 H), 3.09 (m, 2 H),3.45 (m, 2 H), 4.51 (m, 1 H), 4.98 (m, 1 H), 7.09 (m, 4 H), 7.22 (m, 3H), and 7.31 (m, 2 H).

(D) Homophenylalanyl-Nα-Methylornithine 2-(4-Methylphenyl)ethylamide TFA

Titled product (D) is obtained fromBoc-homophenylalanyl-Nγ-Boc-Nα-methylornithine2-(4-methylphenyl)ethylamide, by Procedure E, as a white solid: HPLC(method A); ¹ H NMR (400 MHz, D₂ O) δ 1.61 (m, 2 H), 1.72 (m, 1 H), 1.88(m, 1 H), 2.18 (m, 2 H), 2.30 (s, 3 H), 2.78 (s, 3 H), 2.81 (m, 4 H),3.08 (m, 2 H), 3.58 (m, 2 H), 4.42 (m, 1 H), 4.98 (m, 1 H), 7.18 (m, 4H), 7.40 (m, 3 H), and 7.49 (m, 2 H).

Example 47 Homophenylalanyl-Nα-Methylornithine 2,2-DiphenylethylamideTrifluoroacetate

Titled compound was prepared similarly as in Example 7, except theappropriate homophenyl-alanine precursor was used. FromNα-Boc-N-Fmoc-Nα-methylornithine 2,2-diphenylethylamnide (71 mg), therewas obtained the desired compound (57 mg) as a white solid: ¹ H NMR (400MHz, D₂ O) δ 1.62 (m, 2 H), 1.72 (m, 1 H), 1.89 (m, 1 H), 1.97 (m, 1 H),2.06 (m, 1 H), 2.61 (s, 3 H), 2.79 (m, 1 H), 2.85 (m, 1 H), 3,03 (t,J=7.6 Hz, 2 H), 3.84 (dd, J=13.2; 8.0 Hz, 1 H), 4.15 (dd, J=14.0; 9.2Hz, 1 H), 4.30 (t, J=8.8 Hz, 1 H), 4.38 (m, 1 H), 4.97 (t, J=7.6 Hz, 1H), and 7.52 (m, 15 H).

Example 48 β-(2-Thiazolyl)alanyl-Nα-Methylornithine β-NaphthylamideTrifluoroacetate

(A) Boc-β-(2-Thiazolyl)alanyl-Nγ-Boc-Nα-Methylornithine β-Naphthylamide

Boc-β-(2-thiazolyl)alanine and Nγ-Boc-Nα-methylornithine β-naphthylamidewere coupled under the conditions described in Procedure B, afforded aglassy solid which was purified by silica gel chromatography (1 to 2%MeOH/CH₂ Cl₂): ¹ H NMR (400 MHz, CDCl₃) δ 1.45 (s, 9 H), 1.50 (s, 9 H),1.52 (m, 2 H), 1.77 (m, 1 H), 2.18 (m, 1 H), 3.01 (s, 3 H), 3.19 (m, 3H), 3.49 (m, 1 H), 4.98 (m,1 H), 5.36 (m, 1 H), 7.12 (s, 1 H), 7.43 (m,2 H), 7.59 (d, J=7.2 Hz, 1 H), 7.78 (m, 3 H), 8.28 (s, 1 H), and 8.58(s, 1 H); mass spectrum (ES+) m/e 626 (M+1).

(B) β-(2-Thiazolyl)alanyl-Nα-methylornithine β-NaphthylamideTrifluoroacetate

Deprotection of Boc-β-(2-thiazolyl)alanyl-Nγ-Boc-Nα-methylornithineβ-naphthylamide (A), by Procedure E, afforded a white solid: ¹ H NMR(400 MHz, D₂ O) δ 1.80 (m, 2 H), 1.98 (m, 1 H), 2.19 (m, 1 H), 3.07 (s,3 H), 3.18 (m, 2 H), 3.52 (dd, J=13.0; 6.7 Hz, 1 H), 3.67 (dd, J=13.0;3.5 Hz, 1 H), 5.02 (m, 1 H), 5.29 (t, J=7.9 Hz, 1 H), 7.48 (s, 1 H),7.65 (m, 3 H), 8.03 (m, 3 H), 8.17 (s, 1 H), and 8.88 (s, 1 H); massspectrum (ES+) m/e 426 (M+1).

Example 49 4-(O-Dimethylaminoethyl)tyrosyl-Nα-Methylornithineβ-Naphthylamide Trifluoroacetate

(A) Boc-4-(O-Dimethylaminoethyl)tyrosine

A mixture of Boc-tyrosine, sodium hydride (4 eq), N,N-dimethylaminoethylchloride hydrochloride (1.5 eq), and dimethylformamide (0.1 M) wasstirred at 0° C. for 1 hr. The reaction mixture was then maintained at25° C. (4 h), quenched with water, concentrated in vacuo and flirtherpurified by reverse phase chromatography to give titled compound as asticky solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.39 (s, 9 H), 2.75 (dd,J=17.1; 12.5 Hz, 1 H), 2.89 (s, 6 H), 3.08 (dd, J=12.5; 6.3 Hz, 1 H),3.48 (m, 2 H), 4.19 (m, 1 H), 4.28 (m, 2 H), 6.88 (d, J=9.9 Hz, 2 H),and 7.15 (d, J=9.0 Hz, 2 H).

(B) Boc-4-(O-Dimethylaminoethyl)tyrosyl-Nγ-Boc-Nα-MethylornithineB-Naphthylamide

Using Procedure B, Boc-4-(O-Dimethylaminoethyl)tyrosine (31 mg) andNγ-Boc-Nα-methylornithine β-naphthylamide (43 mg) were condensed toafford a glassy solid (26 mg) after chromatography: ¹ H NMR (400 MHz,CDCl₃) δ 1.42-1.57 (m, 21 H), 1.77 (m, 1 H), 2.79 (m, 4 H), 2.95 (m, 2H), 3.02 (s, 6 H), 3.10 (m, 1 H), 3.61 (m, 2 H), 4.33 (m, 3 H), 4.64 (m,1 H), 6.90 (d, J=8.5 Hz, 2 H), 7.18 (d, J=8.5 Hz, 2 H), 7.41 (m, 3 H),7.74 (m, 3 H), and 8.11 (s, 1 H); mass spectrum (ES+) m/e 706 (M+1).

(C) 4-(O-Dimethylaminoethyl)tyrosyl-Nα-Methylornithine β-NaphthylamideTrifluoro-acetate

Titled product was obtained fromBoc-4-(O-Dimethylaminoethyl)tyrosyl-Nγ-Boc-Nα-methylornithineβ-naphthylamide after exposure to trifluoroacetic acid: HPLC (method A,retention time=35.36 min); ¹ H NMR (400 MHz, D₂ O) δ 1.80 (m, 3 H), 2.07(m, 1 H), 2.69 (s, 9 H), 2.81 (m, 2 H), 3.01-3.29 (m, 3 H), 3.42 (dd,J=11.8; 4.1 Hz, 1 H), 3.57 (m, 1 H), 4.81 (HOD with 3 H hidden), 5.27(t, J=6.4 Hz, 1 H), 6.58 (d, J=9.9 Hz, 2 H), 7.26 (d, J=9.9 Hz, 2 H),7.67 (m, 3 H), 8.08 (m, 3 H), and 8.21 (s, 1 H); mass spectrum (ES+) m/e507 (M+1).

Example 50 4-(O-Methylcarboxyamido)tyrosyl-Nα-Methylornithineβ-Naphthylamide Trifluoroacetate

(A) Boc-4-(O-Methylcarboxyamido)tyrosyl-Nγ-Boc-Nα-methylornithineβ-Naphthyl-amide

A mixture of Boc-tyrosyl-Nγ-Boc-Nα-methylornithine β-naphthylamide (43mg), tetra-butylammonium bromide (5.8 mg), iodoacetamide (14 mg),potassium carbonate (45 mg), and dimethylformamide (0.7 ml) was stirredat 25° C. for 10 hrs. The mixture was then poured into ethyl acetate andworked up to give titled product (47.6 mg) which is used crude: ¹ H NMR(400 MHz, CDCl₃) δ 1.48 (s, 18 H), 1.82 (m, 4 H), 2.79 (s, 3 H),2.82-3.18 (m, 4 H), 4.09 (m, 1 H), 4.50 (m, 2 H), 4.83 (m, 1 H), 6.97(d, J=9.8 Hz, 2 H), 7.21 (d, J=9.8 Hz, 2 H), 7.43 (m, 3 H), 7.79 (m, 3H), and 8.03 (s, 1 H); mass spectrum (ES+) m/e 714 (M+23).

(B) 4-(O-Methylcarboxyamido)tyrosyl-Nα-Methylornithine β-NaphthylamideTFA

Treatment ofBoc-4-(O-methylcarboxyamido)tyrosyl-Nγ-Boc-Nα-methylornithineβ-naphthylamide with trifluoroacetic acid (Procedure E) afforded titledproduct as a white solid; HPLC (method A, retention time=37.34 min); ¹ HNMR (400 MHz, D₂ O) δ 1.79 (m, 3 H), 2.06 (m, 1 H), 2.80 (s, 3 H), 3.12(m, 3 H), 3.41 (dd, J=13.2; 4.4 Hz, 1 H), 3.66 (dd, J=14.8; 2.8 Hz, 1H), 3.83 (d, J=14.8 Hz, 1 H), 4.93 (m, 1 H), 5.22 (t, J=6.8 Hz, 1 H), 6.61 (d, J=7.2 Hz, 2 H), 7.28 (d, J=7.2 Hz, 2 H), 7.55 (d, J=9.2 Hz, 1 H),7.65 (m, 2 H), 7.97 (d, J=8.4 Hz, 1 H), 8.01 (d, J=7.2 Hz, 1 H), and8.06 (m, 2 H); mass spectrum (ES+) m/e 492 (M+1).

Example 51 β-(1-Naphthyl)alanyl-N.sub.α -Methylornithine BenzylamideTrifluoroacetate

(A) Nα-Boc-Nγ-Fmoc-Nα-Methylornithine Benzylamide

This compound is prepared using Procedure C.Nα-Boc-Nγ-Fmoc-Nα-methylornithine (100 mg) was coupled with benzylamineto afford titled compound (50 mg) as a glassy solid: ¹ H NMR (400 MHz,CDCl₃) δ 1.42 (s, 9 H), 1.52 (m, 2 H), 1.70 (m, 2 H), 2.80 (s, 3 H),3.26 (m, 2 H), 4.18 (m, 1 H), 4.22 (m, 1 H), 4.41 (m, 2 H), 4.61 (m, 2H), 7.31 (m, 7 H), 7.43 (m, 2 H), 7.63 (m, 2 H), and 7.81 (m, 2 H).

(B) Boc-β-(l-Naphthyl)alanyl-Nγ-Fmoc-Nα-Methylornithine Benzylamide

This compound is prepared in two steps.Nα-Boc-Nγ-Fmoc-Nα-methylornithine benzylamide (A) (115 mg) isdeprotected with trifluoroacetic acid (5 ml), concentrated andcoevaporated thrice with toluene. The residue is coupled withBoc-β-(1-naphthyl)alanine (87 mg), using Procedure D, to give the titledproduct (45 mg) as a glassy solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.42 (s, 9H), 1.56 (m, 2 H), 1.62 (m, 2 H), 2.59 (s, 3 H), 3.18 (m, 2 H), 3.40(dt, 1 H), 3.59 (dd, 1 H), 4.18 (m, 1 H), 4.21 (m, 1 H), 4.36 (m, 2 H),4.44 (d, 2 H), 7.20-7.40 (m, 12 H), 7.60 (m, 4 H), and 7.79 (m, 4 H).

(C) β-(1-Naphthyl)alanyl-Nα-Methylornithine Benzylamide Trifluoroacetate

Boc-β-(1-Naphthyl)alanyl-Nγ-Fmoc-Nα-methylomithine benzylamide (B) (28mg) was deprotected by Procedure E to afford a white solid: HPLC (methodA); ¹ H NMR (400 MHz, D₂ O) δ 1.59 (m, 2 H), 1.77 (m, 1 H), 1.89 (m, 1H), 2.38 (s, 3 H), 3.03 (t, 2 H), 3.60 (dd, 1 H), 3.82 (dd, 1 H), 4.39(d, 2 H), 4.92 (t, 1 H), 5.01 (dd, 1 H), 7.36-7.50 (m, 12 H), 7.75 (m, 2H), 7.99 (t, 1H), and 8.10 (d, 1 H), and 8.18 (d, 1 H).

Example 52 β-(2-Naphthyl)alanyl-Nα-Methylornithine BenzylamideTrifluoroacetate

This compound was prepared from Nγ-Fmoc-Nα-methylomithine benzylamideand Boc-β-(2-naphthyl)alanine, similar to the procedure in Example 51: ¹H NMR (400 MHz, D₂ O) δ 1.60 (m, 2 H), 1.79 (m, 1 H), 1.98 (m, 1 H),2.82 (s, 3 H), 3.03 (t, 2 H), 3.37 (dd, 1 H), 3.46 (dd, 1 H), 4.22 (s, 2H), 4.97 (m, 2 H), 7.30 (d, 1 H), 7.39 (d, 1 H), 7.43 (m, 3 H), 7.66 (m,2 H), 7.85 (s, 1 H), 7.97 (d, 2 H), and 8.02 (m, 2 H).

Example 53 β-(2-Naphthyl)alanyl-Nα-Methylornithine2-(4-Hydroxyphenyl)ethylamide Trifluoroacetate

(A) Nα-Benzyl-Nγ-Boc-Nα-Methylornithine 2-(4-Hydroxyphenyl)ethylamide

This compound is prepared using Procedure C by coupling ofNα-benzyl-Nγ-Boc-Nα-methylornithine and (4-hydroxyphenyl)ethylamine.

(B) Nγ-Boc-Nα-Methylornithine 2-(4-Hydroxyphenyl)ethylamide

Hydrogen gas was bubbled through a solution ofNα-benzyl-Nγ-Boc-Nα-methylornithine 2-(4-hydroxyphenyl)ethylamide (A)(540 mg) in ethanol (20 ml) in the presence of 1 eq. of conc.hydrochloric acid (1.30 ml) and 5% palladium-on-charcoal (50 mg). Afterdisappearance of starting material, as determined by thin-layerchromatography, the reaction mixture is filtered through a 0.45 μm nylonpad and concentrated in vacuo.

(C) Boc-β-(2-Naphthyl)alanyl-Nγ-Boc-Nα-Methylornithine2-(4-Hydroxyphenyl)ethyl-amide

Using Procedure B, coupling of Boc-β-(2-naphthyl)alanine andNγ-Boc-Nα-methylornithine 2-(4-hydroxyphenyl)ethylamide, followed bysilica gel chromatography (2.5% MeOH/CH₂ Cl₂): ¹ H NMR (400 MHz, CDCl₃)δ 1.41 (m, 21 H), 1.85 (m, 1 H), 2.20 (m, 2 H), 2.45 (m, 1 H), 2.67 (m,4 H), 3.03 (m, 4 H), 4.91 (m, 2 H), 6.77 (m, 2 H), 6.87 (d, J=8.7 Hz, 1H), 6.98 (d, J=10.0 Hz, 1 H), 7.34 (m, 1 H), 7.46 (m, 2 H), 7.66 (m, 1H), and 7.78 (m, 3 H); mass spectrum (ES+) m/e 663 (M+1).

(D) β-(2-Naphthyl)alanyl-Nα-Methylornithine2-(4-Hydroxyphenyl)ethylamide TFA

Boc-β-(2-Naphthyl)alanyl-Nγ-Boc-Nα-methylornithine2-(4-hydroxyphenyl)ethylamide (C) was transformed, by Procedure E, to awhite product; HPLC (method C); ¹ H NMR (400 MHz, CD₃ OD) δ 1.62 (m, 3H), 1.86 (m, 1 H), 2.63 (t, J=7.6 Hz, 2 H), 2.79 (s, 3 H), 2.91 (m, 2H), 3.21 (m, 2 H), 4.71 (t, J=6.0 Hz, 1 H), 4.94 (t, J=6.5 Hz, 1 H),6.69 (d, J=9.4 Hz, 2 H), 7.02 (d, J=9.4 Hz, 2 H), 7.39 (d, J=10.5 Hz, 1H), 7.48 (m, 2 H), 7.78 (s, 1 H), and 7.84 (m, 3 H); mass spectrum (ES+)m/e 463 (M+1).

Example 54 D-Ornithyl-D-β-(2-Naphthyl)alanine BenzylamideTrifluoroacetate

(A) Boc-D-β-(2-Naphthyl)alanine Benzylamide

A mixture of Boc-D-β-(2-naphthyl)alanine (305 mg), benzylamine (165 μL),and ethyl acetate (10 ml) was treated with a solution ofdicyclohexylcarbodiimide (212 mg) in ethyl acetate (5 ml). The mixturewas stirred 3 h at 25° C. and filtered. The mother liquor was dilutedwith ethyl acetate and worked up as usual. The crude residue is used inthe subsequent step: ¹ H NMR (400 MHz, CDCl₃) δ 1.38 (s, 9 H), 3.01 (dd,1 H), 3.34 (dd, 1 H), 4.18 (m, 1 H), 4.38 (t, 1 H), 4.42 (m, 1 H), 7.00(m, 2 H), 7.19 (m, 2 H), 7.33 (m, 2 H), 7.49 (m, 2 H), 7.66 (d, 1 H),and 7.81 (m, 3 H).

(B) Nα,Nγ-Boc-D-Ornithyl-D-β-(2-Naphthyl)alanine Benzylamide

Boc-D-(2-naphthyl)alanine benzylamide (A) (275 mg) is treated withtrifluoroacetic acid (5 ml), concentrated and coevaporated thrice withtoluene. This residue, Nα,Nγ-Boc-ornithine (225 mg),diisopropylethylamine (121 μl), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (135 mg), and methylene chloride (5 ml) was stirred for 3hr at 25° C. The reaction mixture is then poured into ethyl acetate andworked up as usual; the crude product is used in the subsequent step: ¹H NMR (400 MHz, CDCl₃) δ 1.42 (s, 18 H), 1.77 (m, 2 H), 1.98 (m, 2 H),2.97 (m, 1 H), 3.08 (m, 1 H), 3.26 (dd, 1 H), 3.39 (m, 1 H), 4.04 (m, 1H), 4.37 (m, 2 H), 4.79 (q, 1 H), 7.03 (m, 2 H), 7.18 (m, 3 H), 7.38 (d,1 H), 7.49 (m, 2 H), 7.63 (s, 1 H), and 7.79 (m, 3 H).

(C) D-Ornithyl-D-β-(2-Naphthyl)alanine Benzylamide Trifluoroacetate

Treatment of Nα,Nγ-Boc-D-ornithyl-D-β-(2-naphthyl)alanine benzylamide(B) with trifluoro-acetic acid afforded the desired product as a whitesolid: HPLC (method A, retention time=40.43 min); ¹ H NMR (400 MHz, D₂O) δ 1.80 (m, 2 H), 2.02 (m, 2 H), 3.06 (m, 2 H), 3.23 (dd, 1 H), 3.42(dd, 1 H), 4.06 (d, 1 H), 4.17 (t, 1 H), 4.39 (dd, 1 H), 4.81 (HOD withproton hidden), 6.70 (d, 2 H), 6.99 (t, 2 H), 7.18 (t, 1 H), 7.47 (d, 1H), 7.64 (m, 2 H), 7.70 (s, 1 H), 7.93 (m, 2 H), and 8.02 (m, 1 H); massspectrum (ES+) m/e 419 (M+1).

Example 55 D-Ornithyl-D-β-(1-Naphthyl)alanine BenzylamideTrifluoroacetate

(A) Boc-D-β-(1-Naphthyl)alanine Benzylamide

This compound is prepared by coupling of Boc-D-β-(1-naphthyl)alanine andbenzylamine by Procedure C: ¹ H NMR (400 MHz, CDCl₃) δ 1.40 (s, 9 H),3.46-3.61 (m, 2 H), 4.19 (m, 1 H), 4.26 (dd, 1 H), 4.50 (q, 1 H), 7.21(m, 3 H), 7.36 (m, 3 H), 7.49 (m, 1 H), 7.58 (t, 1 H), 7.78 (m, ¹ H),7.85 (t, ¹ H), and 8.20 (d, ¹ H).

(B) Nα,Nγ-Boc-D-Ornithyl-D-β-(1-Naphthyl)alanine Benzylamide

This compound is prepared by coupling D-β-(1-naphthyl)alaninebenzylamide and Nα,Nγ-Boc-D-ornithine, in the presence ofdiisopropylethylamine and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, in methylene chloride: ¹ HNMR (400 MHz, CDCl₃) δ 1.38 (s, 9 H), 1.43 (m, 11 H), 1.64 (m, 2 H),3.02 (m, 1 H), 3.11 (m, ¹ H), 4.06 (m, 2 H), 4.22 (dd, 1 H), 4.33 (dd, 1H), 4.79 (q, 1 H), 7.00 (m, 2 H), 7.22 (m, 3 H), 7.36 (m, 2 H), 7.61 (t,¹ H), 7.68 (t, ¹ H), 7.77 (d, ¹ H), 7.85 (d, 1 H), and 8.21 (d, ¹ H).

(C) D-Ornithyl-D-β-(1-Naphthyl)alanine Benzylamide Trifluoroacetate

The titled product was obtained fromNα,Nγ-Boc-D-ornithyl-D-β-(1-naphthyl)alanine benzyl-amide (B) byProcedure E; HPLC (method A, retention time 37.15 min.); ¹ H NMR (400MHz, D₂ O) δ 1.83 (m, 2 H), 2.05 (m, 2H), 3.10 (m, 2 H), 3.56 (dd, 1 H),3.73 (dd, ¹ H), 4.02 (d, ¹ H), 4.19 (m, 1 H), 6.75 (d, 2 H), 7.30 (m, 2H), 7.45 (t, 1 H), 7.48 (t, 1 H), 7.68 (m, 2 H), 7.87 (d, ¹ H), 8.08 (d,1 H), 8.14 (t, 1 H), and 8.19 (d, 1 H).

Example 56 D-Ornithyl-D-β-(2-Naphthyl)alanine2-(4-Hydroxyphenyl)ethylamide Trifluoroacetate

(A) Boc-D-β-(2-Naphthyl)alanine 2-(4-Hydroxyphenyl)ethylamide

This compound is prepare, by Procedure C, by coupling ofBoc-D-β-(2-naphthyl)alanine with 2-(4-hydroxyphenyl)ethylamine: ¹ H NMR(400 MHz, CD₃ OD) δ 1.36 (s, 9 H), 2.57 (m, 2 H), 2.97 (dd, J=13.8; 11.6Hz, 1 H), 3.18-3.40 (m, 3 H), 4.32 (m, 1 H), 6.67 (d, J=11.4 Hz, 2 H),6.91 (d, J=10.1 Hz, 2 H), 7.34 (d, J=9.5 Hz, 1 H), 7.41 (m, 2 H), 7.66(s, 1 H), and 7.79 (m, 3 H).

(B) Nα,Nγ-Boc-D-Omnithyl-D-β-(2-Naphthyl)alanine2-(4-Hydroxyphenyl)ethylamide

The titled compound is prepared by coupling D-β-(2-naphthyl)alanine2-(4-hydroxyphenyl)ethylamide and Nα,Nγ-Boc-D-ornithine by Procedure C:¹ H NMR (400 MHz, CDCl₃) δ 1.36 (s, 9 H), 1.47 (broad s, 12 H), 1.59 (m,1 H), 2.57 (m, 2 H), 2.97 (m, 2 H), 3.25 (m, 2 H), 3.36 (m, 2 H), 4.00(m, 1 H), 4.72 (m, 1 H), 6.70 (d, J=10 Hz, 2 H), 6.79 (d, J=10 Hz, 2 H),7.30 (d, J=l Hz, 1 H), 7.43 (m, 2 H), 7.62 (s, 1 H), and 7.74 (m, 3 H);mass spectrum (ES+) m/e 647 (M+1).

(C) D-Ornithyl-D-β-(2-Naphthyl)alanine 2-(4-Hydroxyphenyl)ethylamide TFA

The titled product was obtained fromNα,Nγ-Boc-D-ornithyl-D-β-(2-naphthyl)alanine2-(4-hydroxyphenyl)ethylamide, by the Procedure E, as a white solid:HPLC (method C); ¹ H NMR (400 MHz, D₂ O) δ 1.79 (m, 2 H), 1.98 (m, 2 H),2.34 (m, 1 H), 2.42 (m, 1 H), 3.05 (m, 2 H), 3.18 (dd, J=12.9; 11.0 Hz,1 H), 3.26 (dd, J=13.7; 11.0 Hz, 1 H), 3.42 (m, 1 H), 4.07 (t, J=9.0 Hz,1 H), 4.63 (t, J=9.6 Hz, 1 H), 6.72 (d, J=10.0 Hz, 2 H), 6.80 (d, J=10.0Hz, 2 H), 7.44 (d, J=11.5 Hz, 1 H), 7.64 (m, 2 H), 7.80 (s, 1 H), and7.97 (m, 3 H); mass spectrum (ES+) m/e 449 (M+1).

Example 57 D-Ornithyl-D-β-(2-Naphthyl)alanine IsoamylamideTrifluoroacetate

(A) Boc-D-β-(2-Naphthyl)alanine Isoamylamide

Boc-D-β-(2-naphthyl)alanine (190 mg) and isoamylamine were coupled usinga modification of Procedure C to afford titled compound (233 mg) as aglassy solid: ¹ H NMR (400 MHz, CDCl₃) δ 0.89 (t, 6 H), 1.38 (m, 1 H),1.08 (m, 2 H), 1.42 (s, 9 H), 3.18 (m, 3 H), 3.26 (dd, 1 H), 4.39 (q, 1H), 7.38 (d, 1 H), 7.45 (m, 2 H), 7.64 (m, 1 H), and 7.80 (m, 3 H).

(3) Nα,Nγ-Boc-D-Ornithyl-D-β-(2-Naphthyl)alanine Isoamylamide

A solution of Boc-β-(2-naphthyl)alanine benzylamide (225 mg) andtrifluoroacetic acid (8 ml), was stirred for 2 hrs, concentrated andcoevaporated thrice with toluene. This residue is dissolved indichloromethane (10 ml) and Nα,Nγ-Boc-D-ornithine (293 mg),diisopropylethylamine (0.14 ml) and1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (240 mg) added. Afterstirring for 10 hrs, the mixtures was poured into ethyl acetate andworked up as usual. Flash chromatography gave (B) (350 mg) as a glassysolid: ¹ H NMR (400 MHz, CDCl₃) δ 0.80 (t, 6 H), 1.20 (m, 2 H), 1.26 (s,9 H), 1.38 (m, 1 H), 1.41 (s, 9 H), 1.57 (m, 2 H), 1.71 (m, 2 H), 2.97(m, 1 H), 3.10 (m, 2 H), 3.22 (m, 2 H), 3.37 (dd, 1 H), 4.02 (q, 1 H),4.73 (q, 1 H), 7.35 (dd, 1 H), 7.46 (m, 2 H), 7.65 (s, 1 H), and 7.80(m, 3 H).

(C) D-Ornithyl-D-β-(2-Naphthyl)alanine Isoamylamide Trifluoroacetate

Deprotection of Nα,Nγ-Boc-D-ornithyl-D-β-(2-naphthyl)alanineisoamylamide (B) (341 mg), by Procedure E, afforded titled product as awhite solid (308 mg): HPLC (method A, retention time 38.50 min); ¹ H NMR(400 MHz, D₂ O) δ 0.50 (dd, 6 H), 0.91 (m, 3 H), 1.81 (m, 1 H), 2.02 (m,2 H), 2.83 (m, 1 H), 3.09 (m, 3 H), 3.17 (dd, 1 H), 3.30 (dd, 1 H), 4.06(t, 1 H), 4.62 (dd, 1 H), 7.50 (d, 1 H), 7.61 (m, 2 H), 7.79 (m, 1 H),and 7.99 (m, 3 H).

Example 58 D-Ornithyl-N-(Phenethyl)glycine 2-NaphthylamideTrifluoroacetate

(A) Methyl N-(phenethyl)glycinate

A cold solution (0° C.) of glycine methyl ester hydrochloride (1.0 g, 8mmol), methanol (25 mL), glacial acetic acid (0.8 mmol, andphenylacetaldehyde (0.481 g, 4 mmol) was treated with sodiumtriacetoxyborohydride (1.7 g, 8 mmol) in two portions. The reactionmixture was maintained at 0° C. for 1.5 hr, and then quenched withsaturated sodium bicarbonate (15 mL). The solution was extracted withethyl acetate. The organic phase was collected, dried over anhydroussodium sulfate, and adsorbed onto silica gel (100 mg) and applied to acolumn prepacked with silica gel. The column was eluted with CH₂ Cl₂/MeOH (97:3, v:v) to afford titled compound (258 mg).

(B) Methyl N-Boc-N-(phenethyl)glycinate

Methyl N-(phenethyl)glycinate (250 mg, 1.3 mmol) was dissolved in 20 mLof 1:1-water/dioxane, and sodium bicarbonate (2.6 mmol) anddi-tert-butyl dicarbonate (1.9 mmol) were added. After 14 hr at 25° C.,the dioxane was concentrated in vacuo and the aqueous solutionneutralized to pH4 with 5% citric acid (5 mL) and extracted with ethylacetate. The organic phase was collected, dried over anhydrous sodiumsulfate, and concentrated in vacuo to afford titled compound (358 mg): ¹H NMR (400 MHz, CDCl₃) δ 1.44-1.46 (9 H), 2.80-2.89 (2 H), 3.46-3.54 (2H), 3.73 (3 H), 3.77 (1 H), 3.89 (1 H), and 7.0-7.3 (5 H).

(C) N-Boc-N-(Phenethyl)glycine

A solution of methyl N-Boc-N-(phenethyl)glycinate (0.358 g, 1.2 mmol),methanol (20 mL) and 1 M sodium hydroxide (2.4 ml, 2.4 mmol) was stirredfor 14 hr at 25° C. After concentration in vacuo, the residue wasdissolved in water (25 mL) and adjusted to pH4 with 5% citric acid (10mL). The mixture was extracted with ethyl acetate (30 mL) and theorganic phase dried over anhydrous sodium sulfate, and concentrated toafford titled carboxylic acid (311 mg).

(D) Nα,Nγ-Bis-Boc-D-ornithyl-N-(phenethyl)glycine 2-Naphthylamide

A cold solution (0° C.) of Nα,Nγ-Bis-Boc-D-ornithine (130 mg, 0.39mmol), diisopropylethylamine (1.6 mmol) and methylene chloride (10 mL)was treated with PyBroP (275 mg, 0.59 mmol) and kept at 0° C. for 30min. In another reaction, a solution of Boc-N-(phenethyl)glycine2-naphthylamide (240 mg, 0.59 mmol) (made by coupling ofN-Boc-N-(phenethyl)glycine and 2-naphthylamine) and trifluoroacetic acid(3 mL) was stored at 24° C. for 1 hr and then concentrated in vacuo. Theresidue was coevaporated twice with methylene chloride and the resultantsolid resuspended in methylene chloride (10 mL) and treated withdiisopropylethyl-amine (1.2 mmol). The two solutions were mixed andstirred at 25° C. for 1 hr at which time the mixture was washed with 1Mhydrochloric acid (2×25 mL), sat. sodium bicarbonate (25 mL), and brine(25 mL). The organic layer was dried over anhydrous sodium sulfate andadsorbed onto silica gel (500 mg) and applied to a column prepacked withsilica gel. The column was eluted with ethyl acetate:hexane (50:50, v:v)to afford the titled compound.

(E) D-Ornithyl-N-(Phenethyl)glycine 2-Naphthylamide Trifluoroacetate

A solution of Nα,Nγ-BisBoc-D-omithyl-N-(phenethyl)glycine2-naphthylamide (100 mg) and trifluoroacetic acid (10 mL) was maintainedat 25° C. for 1 hr, and then concentrated in vacua. The residue waschromatographed on a reverse-phase column (Amberchrome) with elutionwith acetonitrile/0.1% aqueous trifluoroacetic acid. The appropriatefractions were lyophilized to afford the titled compound (45 mg):

Example 59 Homophenylalanyl-Nα-Methylornithine 3-PhenylpropylamideTrifluoroacetate

A solution of Nα-benzyl-Nγ-Boc-Nα-methylornithine (300 mg),diisopropylethylamine (326 μl), and anhydrous tetrahydrofuran (4 ml) wastreated with PyBop (585 mg) at 0° C. for 10 min., followed by theaddition of a solution of 3-phenylpropylamine (147 μl) in anhydroustetrahydrofuran (2 ml). The resulting solution was stirred at ambienttemperature for 4 hrs. The reaction was diluted with ethyl acetate andwashed with water. The organic layer was dried over anhydrous sodiumsulfate and concentrated in vacuo. The product was purified bychromatography over silica gel (hexane/ethyl acetate) to giveNα-benzyl-Nγ-Boc-Nα-methylornithine 3-phenylpropylamide (370 mg): ¹ HNMR (400 MHz, CDCl₃) δ 1.43 (s, 9 H), 1.6-1.9 (m, 6 H), 2.2 (s, 3 H),2.65 (t, J=6.2 Hz, 2 H), 3.0-3.4 (m, 5 H), 3.64 (s, 2 H), 4.8 (s, 1 H),and 7.1-7.5 (m, 10 H).

A mixture of the above product, methanol (50 ml), 6N hydrochloric acid(141 μl), and 10% palladium-on-carbon (37 mg) was shaken in a Parrhydrogenator (40 psi) for 24 hours. The catalyst was filtered and thefiltrate was concentrated in vacuo. The residue was dissolved in ethylacetate and washed with aqueous sodium bicarbonate, and the organiclayer was dried and concentrated. The product was purified bychromatography over silica gel (methylene chloride/methanol) to giveNγ-Boc-Nα-methylornithine 3-phenylpropylamide.

A cold solution (0° C.) of N-Boc-homophenylalanlne (221 mg),diisopropylethylamine (228 μl), and anhydrous tetrahydrofuran (4 ml) wastreated with PyBrop (398 mg), followed by the addition of a solution ofNγ-Boc-Nα-methylornithine 3-phenylpropyl-amide (228 mg) intetrahydrofuran (3 ml). The resulting solution was stirred forovernight. The precipitate was filtered and the solid was washed withethyl acetate. The filtrate was concentrated in vacuo and the productpurified by silica gel chromatography (hexane/ethyl acetate) to giveN-Boc-homophenylalanyl-Nγ-Boc-Nα-methylornithine 3-phenylpropylamide(260 mg): ¹ H NMR (400 MHz, CDCl₃) δ 1.43 (s, 18 H), 1.7-2.0 (m, 8 H),2.58 (t, J=6.4 Hz, 2 H), 2.6-2.85 (m, 5 H), 3.0-3.4 (m, 4 H), 4.4-4.5(m, 1 H), 4.9-5.0 (m, 1 H), and 7.1-7.3 (m, 10 OH).

The above product was treated with trifluoroacetic acid (2 ml) for 30min. The solution was concentrated in vacuo and the residue was purifiedby reverse phase HPLC (Amberchrome) with acetonitrile/0.1% aqueoustrifluoroacetic acid as the eluent. The desired fraction was lyophilizedto give homophenylalanyl-Nα-methylornithine 3-phenyl-propylamide: ¹H-NMR (400 MHz, D₂ O) δ 1.6-2.0 (m, 8 H), 2.58 (t, J=6.8 Hz, 2 H),2.6-2.8 (m, 5 H), 3.15 (t, J=7.2 Hz, 2 H), 3.30-3.35 (m, 2 H), 4.15-4.20(m, 1 H), 5.05-5.10 (m, 1 H), and 7.05-7.3 (m, 1OH); mass spectrum, m/e425 (M⁺), 290, 264, and 129.

Example 60 Homophenylalanyl-Nα-Methylornithine3-(4-Methylphenyl)propylamide Trifluoroacetate

This was similarly prepared, as described in Example 59, except3-(4-methylphenyl)-propylamine was used as a starting material: ¹ H NMR(400 MHz, D₂ O) δ 1.40-1.75 (m, 6 H), 1.9-2.1 (m, 2 H), 2.2 (s, 3 H),2.48 (t, J=6.8 Hz, 2 H), 2.6-2.8 (m, 5 H), 3.05-3.15 (m, 2 H), 3.22-3.30(m, 2 H), 4.15-4.20 (m, 1 H), 4.95-5.00 (m, 1 H), and 6.9-7.2 (m, 9 H);mass spectru, m/e 439 (M⁺), 290, 278, 261, and 129.

Example 61 Homophenylalanyl-Nα-Methylornithine3-(4-Methoxyphenyl)propylamide Trifluoroacetate

This was similarly prepared, as described in Example 59, except3-(4-methoxyphenyl)-propylamine was used as a starting material: ¹ H NMR(400 MHz, D₂ O) δ 1.6-2.0 (m, 6 H), 2.05-2.15 (m, 2 H), 2.6 (t, J=6.8Hz, 2 H), 2.8 (t, J=7.2 Hz, 2 H), 3.0 (s, 3 H), 3.05 (t, J=7.2 Hz, 2 H),3.15-3.40 (m, 2 H), 3.85 (s, 3 H), 4.60-4.65 (m, 1 H), 4.95-5.05 (m, 1H), 6.93 (d, J=7.2 Hz, 2 H), 7.15 (d, J=7.2 Hz, 2 H), and 7.25-7.45 (m,5 H); mass spectrum, m/e 455 (M⁺), 294, 290, 277, and 129.

Example 62 Homophenylalanyl-Nα-Methylornithine3-(4-fluorophenyl)propylamide Trifluoroacetate

This was similarly prepared, as described in Example 59, except3-(4-fluorophenyl)-propylamine was used as a starting material: ¹ H NMR(400 MHz, D₂ O) δ 1.65-2.0 (m, 6 H), 2.1-2.3 (m, 2 H), 2.6 (t, J=6.8 Hz,2 H), 2.8 (t, J=6.8 Hz, 2 H), 3.0 (s, 3 H), 3.05 (t, J=6.8 Hz, 2 H),3.15-3.40 (m, 2 H), 4.55-4.65 (m, 1 H), 4.95-5.05 (m, 1 H), and7.05-7.45 (m, 9 H); mass spectrum, m/e 443 (M⁺), 313, 290, 282, and 129.

Example 63 Glycyl-Nα-Methylornithine 2-(Cyclohexyl)ethylamideTrifluoroacetate

This was similarly prepared, as described in Example 59, except2-(cyclohexyl)ethylamine and Boc-glycine were used: mass spectrum, m/e313 (M⁺), 295, 356, 239, 186, and 129.

Example 64 Cyclohexylalanyl-Nα-Methylornithine 2-PhenylethylamideTrifluoroacetate

This was similarly prepared, as described in Example 59, except2-phenylethylamine and Boc-p-(cyclohexyl)alanine were used: ¹ H NMR (400MHz, D₂ O) δ 1.2-2.0 (m,17 H), 2.8-3.0 (m, 5 H), 3.0-3.1 (m, 2 H),3.5-3.7 (m, 2 H), 4.4-4.5 (m, 1 H), 4.8-4.9 (m, 1 H), and 7.3-7.5 (m, 5H); mass spectrum, m/e 403 (M), 381, 282, 250, and 129.

Example 65 Leucyl-Nα-Methylornithine 2-Naphthylamide Trifluoroacetate

A solution of Nα-benzyl-Nγ-Boc-Nα-methylornithine (2.13 g, 6.3 mmol),diisopropyl-ethylamine (2.2 ml, 12.6 mmol), and anhydroustetrahydrofuran (20 ml) was treated with PyBop (4 g, 7.6 mmol) at 0° C.for 10 min., followed by the addition of β-amino-naphthalene (1.09 g,7.6 mmol). The resulting solution was stirred at ambient temperatureovernight. The reaction mixture was diluted with ethyl acetate andwashed with water. The organic layer was dried over anhydrous sodiumsulfate and concentrated in vacuo. The product was purified by silicagel chromatography (hexane/ethyl acetate) to affordNα-benzyl-Nγ-Boc-Nα-methylornithine 2-naphthylamide (2 g): ¹ H NMR (400MHz, CDCl₃) δ 1.43 (s, 9 H), 1.6-1.9 (m, 4 H), 2.38 (s, 3 H), 3.2-3.3(m, 2 H), 3.7-3.8 (m, 2 H), 4.7 (s, 1 H), 7.25-7.8 (m, 1 H), and 8.25(s, 1 H).

A mixture of the above product, methanol (60 ml), 6N hydrochloric acid(723 μl, 4.3 mmol), and 10% palladium-on-carbon (200 mg) was shaken inParr hydrogenator (40 psi) for 24 hours. The catalyst was removed byfiltration and the filtrate was concentrated in vacuo. The residue wasdissolved in ethyl acetate and washed with aqueous sodium bicarbonate.The organic layer was dried and concentrated. The product was purifiedby chromatography with CH₂ Cl₂ /CH₃ OH as the eluent to giveNγ-Boc-Nα-methylornithine 2-naphthylamide (1 g): ¹ H NMR (400 MHz,CDCl₃) δ 1.4 (s, 9 H), 1.6-1.8 (m, 2 H), 2.0-2.2 (m, 2 H), 2.45 (s, 3H), 3.1-3.3 (m, 2 H), 4.1-4.3 (m, 1 H), 5.0 (s, 1 H), 7.4-7.5 (m, 2 H),7.7-7.8 (m, 4H), and 8.4 (s, 1 H).

Nγ-Boc-Nα-methylornithine 2-naphthylamide (149 mg, 0.40 mmol),N-Boc-leucine N-hydroxysuccinimide ester (166 mg, 0.5 mmol) anddirnethylformamide (3 ml) was stirred overnight at 70° C. After cooling,the mixture was diluted with ethyl acetate and washed with water. Theorganic layer was dried over anhydrous sodium sulfate and concentrated.The product was purified by chromatography (silica gel-hexane/ethylacetate) to give N-Boc-leucyl-Nγ-Boc-Nα-methylornithine 2-naphthylamide(44 mg): ¹ H NMR (400 MHz, CDCl₃) δ 0.9-1.0 (m, 6 H), 1.4-1.8 (m, 25 H),3.05 (s, 3 H), 3.1-3.2 (m, 2 H), 4.6-4.7 (m, 1 H), 5.1-5.2 (m, 1 H),7.3-7.5 (m, 2 H), 7.7-7.8 (m, 4 H), and 8.1 (s, 1 H).

N-Boc-leucyl-Nγ-Boc-Nα-methylornithine 2-naphthylamide was treated withTFA at 25° C. for 30 min. The solution was concentrated in vacuo and theresidue purified by reverse phase HPLC (Amberchrome--CH₃ CN/0.1% TFA-H₂O). The desired fraction was lyophilized to give titled product: ¹ H NMR(400 MHz, D₂ O) δ 1.0-1.1 (m, 6 H), 1.7-1.9 (m, 5 H), 2.0-2.2 (m, 211),3.1-3.3 (m, 51), 4.6-4.7 (m, 1 H), 5.1-5.2 (m, 1 H), 7.5-7.7 (m, 3 H),and 7.9-8.1 (m. 4 H); mass spectrum, m/e 384 (M⁺), 271, 241, 194, and129.

Example 66 Glycyl-Nα-(Phenethyl)ornithine 3-PhenylpropylamideTrifluoroacetate

A solution of dicyclohexylcarbodiimide (4.68 g, 23 mmol) and ethylacetate (100 ml) was added to a solution of Nα-Fmoc-Nγ-Boc-omithine(10.3 g, 23 mmol) and pentafluorophenol (4.17 g, 23 mmol) in ethylacetate (200 ml). The resulting mixture was stirred at 25° C. for 2hours and the solid formed during reaction was removed by filtration.The filtrate was concentrated in vacuo to give the activated ester as awhite solid. The activated ester (12.7 g, 20 mmol), 3-phenylpropylamine(2.76 g, 20 mmol), and dimethylformamide (100 ml) was stirred at 25° C.for 4 hours. The solution was then treated with piperidine (5 ml) for 1hour at 25° C. and concentrated in vacuo. The residue was dissolved inethyl acetate and washed with aqueous sodium bicarbonate. The organiclayer was dried, concentrated, and purified by chromatography to giveNγ-Boc-ornithine 3-phenylpropylamide (5 g): ¹ H NMR (400 MHz, CD₃ OD) δ1.4 (s, 9 H), 1.45-1.7 (m, 4 H), 1.8-1.9 (m, 2 H), 2.6 (t, J=6.4 Hz, 2H), 3.0-3.1 (m, 211), 3.2-3.3 (m, 2 H), 3.3-3.4 (m, 1 H), and 7.1-7.3(m, 5 H).

A cold solution (0° C.) of the above amine (3 g, 8.6 mmol), in methanol(20 ml), was treated sequentially with acetic acid (345 μl, 6 mmol) andphenylacetaldehyde (1.2 ml, 10 mmol). Then a solution of sodiumcyanoborohydride (2.7 g, 43 mmol) in methanol (10 ml) was added to themixture slowly. The reaction was stirred for additional 30 minutes andthe solvent was removed in vacuo. The residue was dissolved in ethylacetate and washed with aqueous sodium bicarbonate, and purified bychromatography (silica gel, hexane/ethyl acetate) to giveNα-(phenethyl)ornithine 3-phenylpropylamide (2.64 g): ¹ H NMR (400 MHz,CD₃ OD) δ 1.4 (s, 9 H), 1.45-1.8 (m, 6 H), 2.6 (t, J=6.4 Hz, 2 H),2.65-2.8 (m, 4 H), 3.0-3.2 (m, SH), and 7.1-7.3 (m, 10 H).

A cold solution (0° C.) of Boc-glycine (543 mg, 3.1 mmol),diisopropylethylamine (1 ml, 6.2 mmol), and tetrahydrofuiran (10 ml) wastreated with PyBrop (1.45 g, 3.1 mmol), followed by the addition ofNα-phenethyl-Nγ-Boc-ornithine 3-phenylpropylamide (1.28 g, 2.8 mmol) intetrahydrofuran (5 ml). The resulting solution was stirred overnight.The precipitate was filtered and the solid washed with ethyl acetate.The filtrate was concentrated in vacuo and the product purified bychromatography (silica gel--hexane/ethyl acetate) to giveN-Boc-glycyl-Nα-phenethyl-Nγ-Boc-ornithine 3-phenyl-propylamide (1.2 g).

The above product was treated with tnfluoroacetic acid (5 ml) for 30 minand the solution was concentrated in vacuo. The residue was purified byreverse phase HPLC (Amberchrome--CH₃ CN/0.1% TFA-D₂ O) and the desiredfraction was lyophilized to give desired product: ¹ H NMR (400 MHz, D₂O) δ 1.6-2.0 (m, 6 H), 2.5-2.6 (m, 2 H), 2.8-3.0 (m, 4 H), 3.05-3.15 (m,2 H), 3.4-3.5 (m, 2H), 3.7-3.9 (m, 2H1), 4.5-4.6 (m, 1 H), and 7.1-7.4(m, 10 OH); mass spectrum, m/e 411 (M⁺), 393, 354, 336,276, and 219.

Example 67 Glycyl-Nα-(Phenethyl)ornithine 2-NaphthylamideTrifluoroacetate

This was similarly prepared, as described in Example 66, exceptβ-aminonaphthalene was used as the starting material: ¹ H NMR (400 MHz,D₂ O) δ 1.7-2.2 (m, 4 H), 3.0-3.2 (m, 6 H), 3.8-4.0 (m, 2 H), 5.0-5.1(m, 1 H), and 7.3-6.25 (m, 12 H); mass spectrum, m/e 419 (M⁺), 401, 298,276, 258, and 219.

Example 68 Glycyl-Nα-(Phenethyl)ornithine Quinoline-3-amideTrifluoroacetate

This was similarly prepared, as described in Example 66, except3-aminoquinoline was used as the starting material: ¹ H NMR (400 MHz, D₂O) δ 1.7-2.3 (m, 4 H), 3.0-3.3 (m, 4 H), 3.7-3.9 (m, 2 H), 3.9-4.1 (m, 2H), 5.0-5.1 (t, J=6.4 Hz, 1 H), 7.3-7.5 (m, 5 H), 7.8-8.2 (m, 4 H), 8.8(s, 1 H), and 9.1 (s, 1 H); mass spectrum m/e 420 (M⁺), 363, 346, 219,174, and 145.

Example 69 β-Alanyl-Nα-(Phenethyl)ornithine PhenylpropylamideTrifluoroacetate

This was similarly prepared, as described in Example 66, exceptβ-Boc-alanine was used as the acylating reagent: ¹ H NMR (400 MHz, D₂ O)δ 1.6-2.1 (m, 6 H), 2.7 (t, J=6.4 Hz, 2 H), 2.8-3.1 (m, 6 H), 3.25-3.4(m, 4 H), 3.6-3.7 (m, 2 H), 4.6-4.7 (m, 1 H), and 7.3-7.5 (m, 10 H);mass spectrum, m/e 425 (M⁺), 290, 219, 174, and 105.

Example 70 Glycyl-Nα-(2-Hydroxyphenethyl)ornithine 3-PhenylpropylamideTrifluoroacetate

This was similarly prepared, as described in Example 66, except(2-hydroxyphenyl)-acetaldehyde was used in the reductive amination step:¹ H NMR (400 MHz, D₂ O) δ 1.6-2.0 (m, 6 H), 2.6 (t, J=6.4 Hz, 2 H),2.8-3.0 (m, 4 H), 3.05-3.15 (m, 2 H), 3.4-3.6 (m, 2 H), 3.9-4.0 (m, 2H), 4.5-4.6 (m, 1 H), and 6.8-7.3 (m, 9 H); mass spectrum, m/e 427 (M⁺),370, 353, 292, 235, 174, and 190.

Example 71 Glycyl-Nα-(iso-Amyl)ornithine 3-PhenylpropylamideTrifluoroacetate

This was similarly prepared, as described in Example 66, exceptisovaleraldehyde was used in the reductive amination step: ¹ H NMR (400MHz, D₂ O) δ 0.95 (d, J=6.8 Hz, 6 H), 1.5-2.0 (m, 9 H), 2.7 (t, J=6.4Hz, 2 H), 3.0-3.2 (m, 2 H), 3.25-3.45 (m, 4 H), 4.0-4.2 (m, 2 H), 4.6(t, J=6.4 Hz, 1 H), and 7.3-7.5 (m, 5 H); mass spectrum, m/e 377 (M⁺),359, 320, 242, 185, and 140.

Example 72 Glycyl-Nα-(2-Benzo-[b]furanylmethyl)ornithine3-Phenylpropylamide Trifluoroacetate

This was similarly prepared, as described in Example 66, exceptbenzo[b]furan-2-carboxaldehyde was used in the reductive amination step:¹ H NMR (400 MHz, D₂ O) δ 1.5-2.1 (m, 6 H), 2.5 (t, J=6.4 Hz, 2 H),2.8-3.2 (m, 6 H), 4.2-4.4 (m, 2 H), 6.9 (s, 1 H), and 7.2-7.8 (m, 9 H);mass spectrum, m/e 437 (M), 380, 302, 245, and 131.

Example 73 Glycyl-Nα-(3-Quinolinylmethyl)ornithine 3-PhenylpropylamideTrifluoroacetate

This was similarly prepared, as described in Example 66, exceptquinoline-3-carboxaldehyde was used in the reductive amination step: ¹ HNMR (400 MHz, D₂ O) δ 1.2-2.0 (m, 6H), 2.2-2.4 (m, 2 H), 2.6-2.8 (m, 2H), 3.0-3.2 (m, 2 H), 4.1-4.4 (m, 4 H), 4.8-4.9 (m, 1 H), 7.0-7.1 (m, 2H), 7.2-7.4 (m, 3 H), 7.9-8.2 (m, 4 H), 8.9 (s, 1 H), and 9.1 (s, 1 H);mass spectrum, m/e 448 (M⁺), 430, 391, 313, and 256.

Example 74 Glycyl-Nα-(Phenethyl)ornithine 5-IndanylamideTrifluoroacetate

This was similarly prepared, as described in Example 66, except5-aminoindan was used as a starting material: ¹ H NMR (400 MHz, D₂ O) δ1.7-2.3 (m, 6 H), 2.9-3.2 (m, 8 H), 3.7-4.0 (m, 4 H), 5.0 (t, J=6.4 Hz,1 H), and 7.2-7.5 (m, 8 H).

Example 75 Glycyl-Nα-(Phenethyl)lysine 3-PhenylpropylamideTrifluoroacetate

This was similarly prepared, as described in Example 66, exceptNα-Boc-Nγ-Cbz-lysine was used as a starting material: ¹ H NMR (400 MHz,CD₃ OD) δ 1.5-2.1 (m, 8 H), 2.6 (t, J=6.4 Hz, 2 H), 2.8-3.0 (m, 4 H),3.2-3.4 (m, 2 H), 3.5-3.6 (m, 2 H), 3.7-3.9 (m, 2 H), 4.7-4.8 (m, 1 H),and 7.1-7.3 (m, 10 H); mass spectrum, m/e 453 (M⁺), 354, 300, 247, and219.

Example 76 β-Alanyl-Nα-(Phenethyl)lysine 3-PhenylpropylamideTrifluoroacetate

This was similarly prepared, as described in Example 66, exceptNα-Boc-Nγ-Cbz-lysine and p-Boc-alanine were used as starting materials:¹ H NMR (400 MHz, D₂ O) δ 1.3-1.5 (m, 2 H), 1.7-2.0 (m, 6 H), 2.65 (t,J=6.4 Hz, 2 H), 2.8-3.0 (m, 4 H), 3.0-3.1 (m, 2 H), 3.2-3.4 (m, 4 H),3.7-3.9 (m, 2 H), 4.8-4.9 (m, ¹ H), and 7.2-7.5 (m, 10); mass spectrum,m/e 440 (M⁺ +1), 368, 304, 233, and 188.

Example 77 Glycyl-Nα-(Phenethyl)diaminobutyric acid 3-PhenylpropylamideTrifluoroacetate

This was similarly prepared, as described in Example 66, exceptNα-Cbz-Nγ-Boc-L-diaminobutyric acid was used as the starting material: ¹H NMR (400 MHz, D₂ O) δ 1.8-2.0 (m, 2 H), 2.1-2.4 (m, 2 H), 2.7 (t,J=6.4 Hz, 2 H), 2.9-3.1 (m, 4 H), 3.2-3.4 (m, 2 H), 3.6-3.8 (m, 2 H),3.9-4.1 (m, 2 H), 4.7-4.8 (m, 1 H), and 7.2-7.5 (m, 10 H); massspectrum, m/e 397 (M⁺), 379, 276, 242, and 205.

Example 78 β-Alanyl-Nα-(Phenethyl)diaminobutyric acid3-Phenylpropylamide Trifluoroacetate

This was similarly prepared, as described in Example 66, exceptNα-Cbz-Nγ-Boc-L-diaminobutyric acid and P-Boc-alanine were used asstarting materials: ¹ H NMR (400 MHz, D₂ O) δ 1.8-2.0 (m, 2 H), 2.1-2.4(m, 2 H), 2.7 (t, J=6.4 Hz, 2 H), 2.9-3.1 (m, 6 H), 3.2-3.4 (m, 4 H),3.6-3.7 (m, 2 H), 4.7-4.8 (m, 1 H), and 7.2-7.5 (m, 10 H); massspectrum, m/e 411 (M⁺), 393, 340, 276, and 205.

Example 79 4-Aminobutyryl-Nα-(Phenethyl)diaminopropionic acid3-Phenylpropylamide Trifluoroacetate

This was similarly prepared, as described in Example 66, exceptNα-Cbz-Nβ-Boc-L-diaminopropionic acid and Nγ-Boc-aminobutyric acid wereused as starting materials: ¹ H NMR (400 MHz, D₂ O) δ 1.7-1.9 (m, 4 H),2.4-2.6 (m, 4 H), 2.8-3.0 (m, 4 H), 3.1-3.3 (m, 4 H), 3.5-3.7 (m, 2 H),4.2-4.3 (m, 1 H), and 7.2-7.4 (m, 10 H).

Example 80 4-Aminobutyryl-Nα-(Phenethyl)diaminopropionic AcidQuinoline-3-amide Trifluoroacetate

This was similarly prepared as described in Example 66, exceptNα-Cbz-Nβ-Boc-L-diamino-propionic acid, 3-aminoquinoline, andNγ-Boc-aminobutyric acid were used as starting materials: ¹ H NMR (400MHz, D₂ O) δ 1.9-2.1 (m, 2 H), 2.6-2.9 (m, 2 H), 3.0-3.2 (m, 4 H),3.4-3.5 (m, 1 H), 3.8-4.0 (m, 3 H), 4.8-4.9 (m, 1 H), 7.2-7.4 (m, 5 H),8.0 (t, J=7.2 Hz, 1 H), 8.4 (t, J=7.2 Hz, 1 H), 8.3 (t, J=7.2 Hz, 2 H),9.05 (s, 1 H), and 9.4 (s, 1 H); mass spectrum, m/e 420 (M⁺), 402, 335,299, and 270.

Example 81 Acetimidoylglycyl-Nα-(Phenethyl)ornithine 3-PhenylpropylamideTrifluoroacetate

Glycyl-Nα-(phenethyl)omithine 3-phenylpropylamide trifluoroacetate wastreated with ethyl acetimidate in ethanol at pH9 and the product waspurified by HPLC: ¹ H NMR (400 MHz, D₂ O) δ 1.6-2.0 (m, 6 H), 2.3 (s, 3H), 2.7 (t, J=6.4 Hz, 2 H), 2.9-3.1 (m, 4 H), 3.2-3.4 (m, 2 H), 3.5-3.7(m, 2 H), 4.1-4.3 (m, 2 H), 4.7-4.8 (m, 1 H), and 7.2-7.5 (m, 10 H);mass spectrum, m/e 453 (M⁺), 354, 300, 247, and 219.

Example 82 Homophenylalanine N-(3-Aminopropyl)-3-PhenylpropylamideTrifluoroacetate

A mixture of 3-phenylpropylamnine (0.95 g, 7 mmol) and acrylonitnile(0.55 ml, 8.4 mmol) in ethanol (30 ml) was refluxed for 2 hours to giveN-(2-cyanoethyl)-3-phenylpropylamine.

A solution of Boc-homophenylalanine (136 mg, 0.49 mmol),diisopropylethylamine (169 μl mg, 0.97 mmol), and tetrahydrofuran (3 ml)was treated with PyBrop (248 mg, 0.63 mmol) at 0° C.(2-Cyanoethyl)-3-phenylpropylamine (109 mg, 0.58 mmol) intetrahydro-furan (2 ml) was added dropwise and the reaction mixture wasstirred overnight. The solid was filtered and rinsed with ethyl acetate,and the filtrate was concentrated and purified by chromatography to giveBoc-homophenylalanine (2-cyanoethyl)-3-phenyl-propylamide. A mixture ofabove product (200 mg), 10% palladium-on-carbon (20 mg), and methanol(40 ml) was hydrogenated on a Parr hydrogenator (40 psi) overnight. Thecatalyst was removed by filtration through Celite and the filtrate wasconcentrated in vacuo. The residue was then treated with trifluoroaceticacid for 30 min. The solvent was removed in vacuo and product waspurified by reverse phase HPLC to give white solid: ¹ H NMR (400 MHz, D₂O) δ 1.8-2.1 (m, 6 H), 2.5-2.8 (m, 4 H), 3.0-3.3 (m, 4 H), 3.3-3.7 (m, 2H), 4.0-4.1 (m, 1 H), and 7.3-7.5 (m, 10 H).

Example 83 D-(Cyclohexyl)alanineN-(3-Aminopropyl)-3-(Cyclohexyl)propylamide Trifluoroacetate

A suspension of homophenylalanine N-(3-aminopropyl)-3 -phenylpropylamide(100 mg), platinum dioxide (10 mg), 6N hydrochloric acid (0.1 ml), and20 ml of methanol was hydrogenated in a Parr hydrogenator (40 psi) for24 hours. The catalyst was removed by filtration and the productpurified by reverse phase HPLC to give titled compound: ¹ H NMR (400MHz, D₂ O) δ 0.9-1.05 (m, 4 H), 1.1-1.4 (m, 13 H), 1.6-1.8 (m, 12 H),1.9-2.1 (m, 3 H), 3.05-3.15 (m, 2 H), 3.4-3.7 (m, 4 H), and 4.4-4.5 (m,1 H); mass spectrum, m/e 366 (M⁺), 349, 225, 199, and 182.

Example 84 Homophenylalanyl-N-(3-Aminopropyl)aminoethanol 2-NaphthylEther Trifluoroacetate

This was similarly prepared, as described in Example 82, except(2-naphthoxy)ethylamine was used as a starting material: ¹ H NMR (400MHz, D₂ O) δ 1.9-2.0 (m, 2 H), 2.2-2.4 (m, 2 H), 2.6-2.8 (m, 2 H),2.9-3.1 (m, 2 H), 3.4-3.6 (m, 2 H), 3.6-3.8 (m, 2 H), 4.2-4.4 (m, 2 H),4.5-4.6 (m, 1 H), and 6.9-7.9 (m, 12 H).

Example 85 Homophenylalanyl-Ornithinethiol 2-Phenethyl ThioetherTrifluoroacetate

A well, stirred cold solution (0° C.) of Nα-Boc-Nγ-Cbz-ornithine (5 g,13.6 mmol), diiso-propylethylamine (5.9 ml, 34 mmol), andtetrahydrofuran (60 ml) was treated with ethyl chloro-formate (3.25 ml,34 mmol) at 0° C. Sodium borohydride (2.58 g, 68 mmol) was added,followed by the slow addition of 1:1-tetrahydrofiiran/water (10 ml) 30minutes later. The reaction mixture was acidified with 6N hydrochloricacid and the solution was extracted with dichloromethane. The organiclayer was dried over anhydrous sodium sulfate and purified bychromatography to give Nα-Boc-Nγ-Cbz-ornithinol (3.9 g): ¹ H NMR (400MHz, CDCl₃) δ 1.2-1.4 (m, 13 H), 3.0 (t, J=6.4 Hz, 2 H), 3.4 (s, 2 H),3.8 (s, 1 H), 4.95 (s, 2 H), and 7.1-7.2 (m, 5 H).

Diethyl azodicarboxylate (180 μl, 1.1 mmol) was added to a solution oftriphenylphosphine (288 mg, 1.1 mol) in tetrahydrofirran (3 ml) at 0° C.and the resulting mixture was further stirred for 30 min. at 0° C. Asolution of Nα-Boc-Nγ-Cbz-ornithinol (200 mg, 0.57 mmol), thioaceticacid (86 μl, 1.1 mmol), and tetrahydrofuran (2 ml) was added and themixture was then stirred at 25° C. for 3 hours. The reaction solutionwas diluted with ethyl acetate and washed with aqueous sodiumbicarbonate. The product was purified by chromatography. to give N.sub.α-Boc-N.sub.δ -Cbz-ornithinethiol S-acetate (189 mg) which intetrahydrofuran (3 ml) was reacted with 0.5N sodium methoxide (2 ml) at25° C. for 4 hours. The resulting solution was treated with(2-iodoethyl)benzene (320 mg, 1.4 mmol) and was stirred for overnight.The reaction solution was diluted with ethyl acetate, washed with water,and purified by chromatography to give Nα-Boc-Nγ-Cbz-ornithinethiol2-phenethyl thioether (160 mg): ¹ H NMR (400 MHz, CDCl₃) 5 1.4-1.7 (m,13 H, 2.6-2.9 (m, 6 H), 3.1-3.3 (m, 2 H), 3.7 (s, 1 H), 5.1 (s, 2 H),and 7.2-7.4 (m, 10 H).

The above product was treated with trifluoroacetic acid to remove theBoc protection group, followed by acylation with Boc-homophenylalaninemediated by PyBrop to give Boc-homophenylalanyl-Nγ-Cbz-ornithinethiol2-phenethyl thioether. The protecting groups were removedsequentially - 1) catalytic hydrogenation and 2) trifluoroacetic acid togive homophenylalanyl-ornithinethiol 2-phenethyl thioethertrifluoroacetate: ¹ H NMR (400 MHz, DMSO) δ 1.3-1.6 (m, 4 H), 1.9-2.0(m, 2 H), 2.6-2.8 (m, 10 H), 3.7-3.9 (m, 2 H), and 7.1-7.3 (m, 10 H);mass spectrum, m/e 400 (M⁺), 383, 319, 239, 222, and 200.

Example 86 Phenylalanyl-Ornithinethiol 2-Naphthyl ThioetherTrifluoroacetate

(A) Nα-Boc-Nγ-Cbz-Ornithinol Methanesulfonate

A solution of Nα-Boc-Nγ-Cbz-ornithinol (363 mg) in dichloromethane (10ml) at 0° C., under nitrogen atmosphere, was sequentially treated withmethanesulfonyl chloride (110 μl, 1.4 eq) and triethylamine (200 μl, 1.4eq). After 3 hrs, the solution was poured into dichloromethane andworked up as usual. The crude product, which is very pure (400 mg), is awhite solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.45 (s, 9 H), 1.63 (m, 4 H),3.02 (s, 3 H), 3.24 (m, 2 H), 3.84 (m, 1 H), 4.19 (dd, J=9.5; 3.7 Hz, 1H), 4.23 (d, J=9.5 Hz, 1 H), 5.14 (s, 2 H), and 7.39 (m, 5 H).

(B) Nα-Boc-Nγ-Cbz-ornithinethiol 2-Naphthyl Thioether

A solution of Nα-Boc-Nγ-Cbz-ornithinol methanesulfonate (A) (100 mg) indimethylformamide (2.5 ml), is added sodium iodide (71 mg),β-naphthylthiol (66 mg) and diisopropylethylamine (85 μl) was maintainedat 70° C. for 12 h. After cooling to room temperature, the reactionmixture was poured into ethyl acetate and worked up as usual. Afterchromatography (10 to 30% ethyl acetate/hexane), there was obtainedintermediate (B) (60 mg) as a white solid: ¹ H NMR (400 MHz, CDCl₃) δ1.41 (s, 9 H), 1.42-1.78 (m, 4 H), 3.19 (m, 4 H), 3.86 (m, 1 H), 5.13(s, 2 H), 7.38 (m, 5 H), 7.44 (m, 3 H), 7.75 (d, J=11.3 Hz, 2 H), 7.80(d, J=10.1 Hz, 1 H), and 7.85 (s, 1 H).

(C) Boc-Phenylalanyl-Nγ-Cbz-ornithinethiol 2-Naphthyl Thioether

A solution of Nα-Boc-Nγ-Cbz-ornithinethiol 2-naphthyl thioether (B) (60mg) and 4M hydrochloric acid/dioxane (3 ml) was stirred at 25° C. for1.5 h and then concentrated in vacuo. The crude residue was coupled toBoc-phenylalanine by Procedure C, followed by silica gel chromatography(20 to 30%-ethyl acetate/hexane) to give a glassy solid (73 mg).

(D) Phenylalanyl-Ornithinethiol 2-Naphthyl Trifluoroacetate

A solution of Boc-phenylalanyl-Nγ-Cbz-ornithinethiol 2-naphthylthioether (C) (30 mg) and a mixture of trifluoroaceticacid/triethylsilane (3:1) (10 ml) was stirred at 25° C. for 1 hr andconcentrated in vacuo. The crude material was purified by HPLC to afforddesired product (10 mg) as a white solid: HPLC (method A, retentiontime=46.26 min); ¹ H NMR (400 MHz, D₂ O) δ 1.69 (m, 2 H), 1.77 (m, 1 H),1.87 (m, 1 H), 2.84 (t, J=8.4, 2 H), 3.02 (m, 2 H), 3.09 (dd, J=14.4;7.2 Hz, 1 H), 3.22 (dd, J=14.0; 5.2 Hz, 1 H), 4.15 (t, J=8.0 Hz, 1 H),7.16 (m, 2 H), 7.34 (m, 3 H), 7.48 (m, 1 H), 7.60 (m, 3 H), 7.93 (t,J=7.2, 1 H), and 7.97 (m, 2 H).

Example 87 Homophenylalanyl-Ornithinethiol 2-Benzothiazolyl ThioetherTrifluoroacetate

(A) Nα-Boc-Nγ-Cbz-Ornithinethiol 2-Benzothiazolyl Thioether

A solution of Nα-Boc-Nγ-Cbz-ornithinol methanesulfonate (100 mg), sodiumiodide (70 mg), dimethylformamide (2.5 ml), 2-mercaptobenzothiazole (70mg) and diisopropyl-ethylamine (85 μl) was stirred at 70° C. for 12 h.After cooling to ambient temperature, the reaction mixture was pouredinto ethyl acetate and worked up. The crude material was chromatographedover silica gel (20 to 50% ethyl acetate/hexane) to afford titledproduct (50 mg) as a white solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.39 (s, 9H), 1.58-1.69 (m, 4 H), 3.22 (m, 2 H), 3.54 (m, 2 H), 3.99 (m, 1 H),5.12 (s, 2 H), 7.23-7.42 (m, 7 H), 7.77 (d, J=10.3 Hz, 1 H), and 7.86(d, J=9.7 Hz, 1 H).

(B) Boc-Homophenylalanyl-Nγ-Cbz-Ornithinethiol 2-BenzothiazolylThioether

Coupling of Nγ-Cbz-ornithinol 2-benzothiazolyl thioether andBoc-homophenylalanine afforded a glassy solid: ¹ H NMR (400 MHz, CDCl₃)δ 1.42 (s, 9 H), 1.63-1.79 (m, 5 H), 1.92 (m, 1 H), 2.49 (m, 2 H), 3.23(m, 2 H), 3.58 (broad d, J=14.1 Hz, 1 H), 3.70 (dd, J=13.5; 10.3 Hz, 1H), 4.01 (m, 1 H), 4.30 (m, 1 H), 5.12 (s, 2 H), 6.97 (d, J=9.0 Hz, 1H), 7.18 (m, 2 H), 7.29 (m, 5 H), 7.37 (m, 3 H), 7.48 (t, J=8.3 Hz, 11H), 7.73 (d, J=8.4 Hz, 1 H), and 8.02 (m, 1 H).

(C) Homophenylalanyl-Ornithinethiol 2-Benzothiazolyl ThioetherTrifluoroacetate

A solution of Boc-homophenylalanyl-Nγ-Cbz-ornithinethiol2-benzothiazolyl thioether (B) (35 mg) and trifluoroacetic acid (10 ml)was kept at 25° C. for 2 hrs and concentrated in vacuo. After HPLCpurification (method A, retention time=42.1 min), there was obtained awhite solid (33 mg): ¹ H NMR (400 MHz, D₂ O) δ 1.78-1.90 (m, 4 H), 2.04(m, 2 H), 2.62 (m, 2 H), 3.11 (m, 2 H), 3.44 (dd, J=13.3; 9.6 Hz, 1 H),3.79 (dd, J=13.0; 2.7 Hz, 1 H), 4.06 (t, J=5.9 Hz, 1 H), 4.41 (m, 1 H),7.03 (m, 2 H), 7.24 (m, 3 H), 7.42 (t, J=8.5 Hz, 1 H), 7.55 (t, J=9.4Hz, 1 H), 7.81 (d, J=9.5 Hz, 1 H), and 7.85 (d, J=8.6 Hz, 1 H).

Example 88 D-Phenylalanyl-Ornithinethiol 2-Benzothiazolyl ThioetherTrifluoroacetate

(A) Boc-D-Phenylalanyl-Nγ-CBz-Omithinethiol Benzothiazolyl Thioether

This compound is prepared in two steps. Nα-Boc-Nγ-Cbz-ornithinethiol2-benzothiazolyl thioether (140 mg) is treated with 4M hydrochloric acidin dioxane (5 ml) for 20 min and concentrated in vacuo. The intermediateNγ-CBz-omithinethiol 2-benzothiazolyl thioether is dissolved indimethylformamide (3 ml), diisopropylethylamine (50 μl) andBoc-phenylalanine N-hydroxysucciniide ester are added. After stirring 12h, concentration and sillica gel chromatography (1 to 2%methanol/dichloromethane) afforded pure intermediate (129 mg): ¹ H NMR(400 MHz, CDCl₃) δ 1.43 (s, 9 H), 1.53 (m, 2H), 1.62 (m, 2 H), 2.73 (m,1 H), 2.98 (dd, J=14.2; 7.7 Hz, 1 H), 3.20 (m, 2 H), 3.42 (m, 2 H), 4.23(m, 2 H), 6.12 (s, 2 H), 6.91 (m, 2 H), 7.09 (m, 3 H), 7.35 (m, 6 H),7.44 (t, J=10.6 Hz, 1 H), 7.78 (d, J=9.1 Hz, 1 H), and 7.90 (d, J=10.1Hz, 1 H).

(B) D-Phenylalanyl-Ornithinethiol 2-Benzothiazolyl ThioetherTrifluoroacetate

Boc-D-phenylalanyl-Nγ-CBz-ornithinethiol 2-benzothiazolyl thioether (A)was stirred at 25° C. with trifluoroacetic acid-triethylsilane (3:1-10ml) for 2 hrs and then concentrated in vacuo. The crude residue waspurified by HPLC (method C, retention time=49.57 min); ¹ H NMR (400 MHz,D₂ O) δ 1.46 (m, 3 H), 1.74 (m, 1 H), 2.98 (m, 2 H), 3.22 (m, 2 H), 3.40(dd, J=13.6; 8.0 Hz, 1 H), 3.66 (d, J=14.0 Hz, 1 H), 4.17 (m, 2 H), 7.30(m, 2 H), 7.44 (m, 3 H), 7.52 (t, J=8.0 Hz, 1 H), 7.62 (t, J=7.2 Hz, 1H), 7.94 (d, J=8.4 Hz, 1 H), and 8.02 (d, J=8.4 Hz, 1 H).

Example 89 4-Fluorophenylalanyl-Ornithinethiol 2-BenzimidazolylThioether Trifluoroacetate

This was similarly prepared, as described in Example 87, exceptBoc-4-fluorophenylalanine and 2-mercaptobenzimidazole were used asstarting materials.

Example 90 D-Ornithyl-D-phenylalaninethiol 2-Naphthyl ThioetherTrifluoroacetate

(A) N-Boc-D-Phenylalaninol Methanesulfonate

A cold solution (0° C.) of N-Boc-D-phenylalaninol (0.53 g, 2.09 mmol) inanhydrous methylene chloride (20 mL) was treated sequentially withmethanesulfonyl chloride (360 mg, 3.14 mmol) and triethylamine (0.32 g,3.14 mmol). The reaction was stirred at 0° C. for 2 hr, quenched with 1Mhydrochloric acid (2×25 mL) and extracted with methylene chloride. Thecombined extract was washed with saturated sodium bicarbonate (1×25 mL),and brine (1×25 mL). The organic layer was dried over anhydrous sodiumsulfate, filtered and the filtrate adsorbed onto silica gel and appliedto a column prepacked with silica gel. The title compound was elutedfrom the column with hexane:ethyl acetate (60:40, v:v) to furnish titledcompound (352 mg) as a white solid.

(B) N-Boc-D-phenylalaninethiol 2-Naphthyl Thioether

A mixture of N-Boc-D-phenylalaninol mesylate (0.3 g, 0.9 mmol),dimethylformamide (10 mL), 2-napthalenethiol (0.22 g, 1.39 mmol),diisopropylethylamine (1.39 mmol) and sodium iodide (0.14 g, 0.93 mmol)was kept at 80 ° C. for 18 hr, cooled to room temperature and methylenechloride (25 ml) was added. This mixture was washed with water (2×10mL), 1M sodium hydroxide (3×10 mL) and brine (2×15 mL). The organiclayer was dried over anhydrous sodium sulfate, filtered and the filtrateadsorbed onto silica gel and applied to a column prepacked with silicagel. The title compound (121 mg) was eluted with hexane:ethyl acetate(90:10, v:v) to afford a white solid.

(C) D-Ornithyl-D-phenylalaninethiol 2-Naphthyl ThioetherTrifluoroacetate

N-Boc-D-phenylalaninethiol 2-naphthyl thioether was deprotected(Procedure E) to afford D-phenylalaninethiol 2-naphthyl thioethertrifluoroacetate which was coupled (Procedure B) withNα,Nγ-Bis-Boc-ornithine. The resultantNα,Nγ-bis-Boc-ornithyl-D-phenylalaninethiol 2-naphthyl thioether wasdeprotected by exposure to trifluoroacetic acid to afford after HPLCpurification the titled compound: ¹ H NMR (400 MHz, D₂ O) δ 1.60-1.70 (4H), 2.64-2.70 (2 H), 2.90-3.30 (3 H), 3.41-3.50 (1 H), 3.80-3.82 (1 H),4.39-4.41 (1 H), 7.23-7.70 (8 H), and 7.80-8.05 (4 H); mass spectrum(relative intensity) m/e 408 (100, M+1).

Example 91 D-Lysyl-D-Leucinethiol 2-Benzothiazolyl ThioetherTrifluoroacetate

This was similarly prepared, as described in Example 90, exceptNα,N-bis-Boc-D-lysine and D-leucinethiol 2-benzothiazolyl thioether wereused.

Example 92 D-(3-Chlorotyrosyl)-D-Phenylalaninethiol 2-BenzimidazolylThioether Trifluoroacetate

This was similarly prepared, as described in Example 90, exceptBoc-D-3-chlorotyrosine and D-phenylalaninethiol 2-benzimidazolylthioether were used.

Example 93 D-β-(4-Pyridyl)alanyl-D-Methioninethiol 3,4-DimethoxyphenylThioether Trifluoroacetate

This was similarly prepared, as described in Example 90, exceptBoc-D-β-(4-pyridyl)-alanine and D-methioninethiol 3,4-dimethoxyphenylthioether were used.

Example 94 D-Ornithyl-D-Cysteinethiol 2-Benzimidazolyl ThioetherTrifluoroacetate

This was similarly prepared, as described in Example 90, exceptNα,Nγ-bis-Boc-D-ornithine and D-cysteinethiol 2-benzimidazolyl thioetherwere used.

Example 95 Homophenylalanyl-Ornithinol 2-Naphthyl Ether Trifluoroacetate

(A) Nα-Boc-Nγ-Cbz-Ornithinol

This was prepared from Nα-Boc-Nγ-Cbz-ornithine using Procedure F: ¹ HNMR (400 MHz, CDCl₃) δ 1.43 (broad s, 10 H), 1.57 (m, 3 H), 3.21 (m, 2H), 3.53-3.66 (m, 3 H), 5.09 (s, 2 H), and 7.38 (m, 5 H).

(B) Nα-Boc-Nγ-Cbz-Ornithinol 2-Naphthyl Ether

A solution of Nα-Boc-Nγ-Cbz-ornithinol (587 mg), dichloromethane (17ml), 2-naphthol (288 mg), triphenylphosphine (524 mg) andN,N'-diisopropylazodicarboxamide (393 μl) was stirred for 12 hrs at 25°C., under nitrogen. The reaction mixture was poured into dichloromethaneand washed successively with saturated sodium bicarbonate and brine. Theorganic phase was then dried over anhydrous sodium sulfate andconcentrated in vacuo. Further purification by flash chromatography gavethe titled product (534 mg) as a glassy solid: ¹ H NMR (400 MHz, CDCl₃)δ 1.41 (s, 9 H), 1.60-1.73 (m, 4 H), 3.20 (m, 2 H), 4.01 (m, 3 H), 5.07(s, 2 H), 7.09 (s, 1 H), 7.11 (d, J=8.3 Hz, 1 H), 7.28 (m, 6 H), 7.40(t, J=8.4 Hz, 1 H), and 7.71 (m, 3 H).

(C) Boc-Homophenylalanyl-Nγ-Cbz-Ornithinol 2-Naphthyl Ether

A solution of Nα-Boc-Nγ-Cbz-ornithinol 2-naphthyl ether (273 mg) and 4Mhydrochloric acid/dioxane (3 ml) was stirred at room temperature for 1.5h and then concentrated in vacuo. The crude residue is coupled toBoc-homophenylalanine as per Procedure C, followed by flashchromatography (40% ethyl acetate/hexane) to give desired product (130mg) as a glassy solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.44 (s, 9 H), 1.62(m, 2 H), 1.75 (m, 2 H), 1.91 (m, 1 H), 2.18 (m, 1 H), 2.68 (m, 2 H),3.25 (m, 2 H), 4.05 (m, 2 H), 4.37 (m, 1 H), 5.11 (s, 11 H), 7.09-7.19(m, 7 H), 7.32 (m, 6 H), 7.45 (t, J=9.0 Hz, 1 H), and 7.75 (m, 3 H).

(D) Homophenylalanyl-Ornithinol 2-Naphthyl Ether Trifluoroacetate

Hydrogen was bubbled through a solution of Nα-Boc-Nγ-Cbz-ornithinol2-naphthyl ether (C) (130 mg) and methanol (10 ml), with 10%palladium-on-charcoal (10 mg), until starting material was absent (bythin-layer chromatography). The reaction mixture is filtered through a0.45 μm nylon pad and concentrated in vacuo, and the residue dissolvedin trifluoroacetic acid (2 ml). After 1 hr, the solution wasconcentrated in vacuo: ¹ H NMR (400 MHz, D₂ O) δ 1.82-1.91 (m, 4 H),2.16 (m, 2 H), 2.63 (m, 2 H), 3.17 (m, 2 H), 4.12 (t, J=5.0; 8.0 Hz, 1H), 4.22 (dd, J=11.6; 8.5 Hz, 1 H), 4.37 (dd, J=11.6; 2.3 Hz, 1 H), 4.98(m 1 H), 6.90 (d, J=10.2 Hz, 2 H), 7.07 (t, J=9.0 Hz, 2 H), 7.18 (d,J=7.7 Hz, 1 H), 7.21 (d, J=9.0 Hz, 1 H), 7.40 (s, 1 H), 7.50 (t, J=8.0Hz, 1 H), 7.59 (t, J=7.5 Hz, 1 H), and 7.80-7.92 (m, 3 H).

Example 96 2-Methyltyrosyl-Ornithinol 1-Naphthyl Ether Trifluoroacetate

This was similarly prepared as described in Example 95, exceptBoc-2-methyltyrosine and ornithinol 1-naphthyl ether were used.

Example 97 B-(2-Thienyl)alanyl-Lysinol 3,4-Dimethylphenyl EtherTrifluoroacetate

This was similarly prepared as described in Example 95, exceptBoc-b-(2-thienyl)alanine and lysinol 3,4-dimethylphenyl ether were used.

Example 98 Leucyl-D-Leucinol 2-Benzimidazolyl Ether Trifluoroacetate

This was similarly prepared as described in Example 95, exceptBoc-leucine and leucinol 2-benzimidazolyl ether were used.

Example 99 D-Lysyl-D-Leucinol 3-Quinolinyl Ether Trifluoroacetate

This was similarly prepared as described in Example 95, exceptNα,Nε-bis-Boc-lysine and D-leucinol 3-quinolinyl ether were used.

Example 100 D-Ornithyl-D-Phenylalaninol 2-Naphthyl EtherTrifluoroacetate

This was similarly prepared as described in Example 95, exceptNα,Nγ-bis-Boc-ornithine and D-phenylalaninol 2-naphthyl ether used: ¹ HNMR (400 MHz, D₂ O) δ 2.58-2.80 (4 H), 2.65-2.68 (2 H), 2.98-3.23 (2 H),3.90-4.00 (1 H), 4.20-4.40 (2 H), 4.61-4.64 (1 H), 7.25-7.60 (10 H), and7.80-8.05 (2 H); mass spectrum (relative intensity) m/e 392 (80, M+1).

Example 101 Phenylalanyl-Ornithinol 2-Naphthyl Ether Trifluoroacetate

This was similarly prepared, as described in Example 95, exceptphenylalanine was introduced: ¹ H NMR (400 MHz, D₂ O) δ 1.78-1.89 (m, 4H), 3.10 (m, 3 H), 3.23 (dd, J=13.6; 6.0 Hz, 1 H), 3.88 (dd, J=10.0; 3.6Hz, 1 H), 4.02 (dd, J=10.4; 5.2 Hz, 1 H), 4.27 (m, 2 H), 6.96 (t, J=7.2Hz, 1 H), 7.14 (m, 2 H), 7.23 (m, 3 H), 7.31 (s, 1 H), 7.53 (t, J=8.0Hz, 1 H), 7.64 (t, J=6.8 Hz, 1 H), and 7.97 (m, 3 H).

Example 102 Homophenylalanyl-Nα-Methylornithinol 2-Naphthyl EtherTrifluoroacetate

(A) Nα-Benzyl-Nγ-Boc-Ornithine

A solution of Nγ-Boc-ornithine (7.0 g), 2M sodium hydroxide (20 ml),benzaldehyde (3.2 ml) and methanol (10 ml) was cooled to 0° C. andsodium borohydride (2.7 g) is added. After 1 hr at 0° C., the mixturewas kept at 25° C. for 12 h. Water (100 ml) was added and the mixtureextracted with ether (2×60 ml). The combined organic extract was washedwith sat. sodium bicarbonate (ca. 150 ml) and water, and dried overanhydrous sodium sulfate. After concentration in vacuo, the desiredproduct (4.5 g) was obtained as a white solid: ¹ H NMR (400 MHz, CD₃ OD)δ 1.42 (s, 9 H), 1.60 (m, 2 H), 1.84 (m, 2 H), 3.03 (m, 2 H), 3.50 (t,J=7.4 Hz, 1 H), 4.11 (d, J=11.9 Hz, 1 H), 4.21 (d, J=l 1.9 Hz, 1 H),7.41 (m, 3 H), and 7.49 (m, 2 H).

(B) Nα-Benzyl-Nγ-Boc-Nα-Methylornithine

36% Formalin (5.8 ml) is added to a suspension ofNα-benzyl-Nγ-Boc-ornithine (A) (3.66 g) in acetonitrile (220 ml),methanol (110 ml) and water (110 ml), and the mixture stirred at 25° C.until clear. After cooling to 0° C., sodium cyanoborohydride (1.6 g) wasadded and the mixture maintained at 25° C. for 10 hrs. Water (190 ml)was added, and the mixture acidified with 5% citric acid to pH 3.5.After extracting with chloroform (3×60 ml), the combined organic phaseis washed with brine and dried over anhydrous sodium sulfate. Removal ofthe solvent in vacuo afforded amino acid (2.1 g) as a white solid: ¹ HNMR (400 MHz, CD₃ OD) δ 1.42 (s 9 H), 1.61 (m, 1 H), 1.73 (m, 1 H), 1.97(m, 2 H), 2.77 (s, 3 H), 3.09 (m, 2 H), 3.61 (m, 1 H), 4.30 (m, 2 H),7.46 (m, 3 H), and 7.56 (m, 2 H).

(C) Nα-Benzyl-Nγ-Boc-Nα-Methylornithinol

Using Procedure F, Nα-benzyl-Nγ-Boc-Nα-methylornithine (B) (2.7 g) isconverted to alcohol (C) (2.0 g) as a white solid, after silica gelchromatography (5% methanol/dichloromethane): ¹ H NMR (400 MHz, CDCl₃) δ1.20 (m, 1 H), 1.42 (broad s, 11 H), 1.61 (m, 1 H), 2.18 (s, 3 H), 2.79(m, 1 H), 3.12 (m, 2 H), 3.33 (broad s, 1 H), 3.37 (t, J=10.4 Hz, 1 H),3.52 (m, 2 H), 3.69 (d, J=13.2 Hz, 1 H), and 7.24 (m, 5 H).

(D) Nα-Benzyl-Nγ-Boc-Nα-Methylornithinol 2-Naphthyl Ether

Nα-Benzyl-Nγ-Boc-Nα-methylornithinol (168 mg), dichloromethane (10 ml),2-naphthol (91 mg), triphenylphosphine (165 mg), andN,N'-diisopropylazodicarboxamide (124 μl) was stirred for 12 hrs at 25°C., under nitrogen. The reaction mixture was poured into dichloromethaneand washed successively with saturated sodium bicarbonate and brine. Theorganic phase was then dried over anhydrous sodium sulfate andconcentrated in vacuo. Further purification by flash chromatography gavethe titled product (90 mg, 39%) as a white solid. ¹ H NMR (400 MHz,CDCl₃) δ 1.51 (s, 9 H), 1.68 (m, 2 H), 1.75 (m, 2 H), 2.39 (s, 3 H),3.20 (m, 2 H), 3.82 (d, J=13.3, 11 H), 3.93 (d, J=13.2, 11 H), 4.11 (dd,J=9.6; 3.6, 11 H), 4.29 (dd, J=10.2; 8.4, 1 H), 7.22 (m, 2 H), 7.29 (t,J=7.8, 1 H), 7.39 (m, 5 H), 7.50 (t, J=8.0, 1 H), 7.79 (m, 3 H).

(E) Nγ-Boc-Nα-Methylornithinol 2-Naphthyl Ether

A methanolic solution of Nα-benzyl-Nγ-Boc-Nα-methylornithinol 2-naphthylether (D) was reduced with hydrogen, over 5% palladium-on-carbon, toafford the titled product: ¹ H NMR (400 MHz, CDCl₃) δ 1.44 (s, 9 H),1.63 (m, 2 H), 1.95 (m, 2 H), 2.53 (s, 3 H), 2.95 (m, 1 H), 3.09 (m, 2H), 4.01 (dd, J=10.7; 6.7 Hz, 1 H), 4.12 (dd, J=10.5; 4.4 Hz, 1 H), 7.19(m, 2 H), 7.35 (t, J=9.1 Hz, 1 H), 7.45 (t, J=8.9 Hz, 1 H), and 7.75 (m,3 H).

(F) Boc-Homophenylalanyl-Nγ-Boc-Nα-Methylornithinol 2-Naphthyl Ether

Using Procedure D, coupling of Nγ-Boc-Nα-methylornithinol 2-naphthylether (E) and Boc-homophenylalanine afforded titled compound as a glassysolid: ¹ H NMR (400 MHz, CDCl₃) δ 1.44 (2s, 18 H), 1.58-1.70 (m, 4 H),1.98 (m, 2 H), 2.73 (m, 2 H), 2.83 (s, 3 H), 3.18 (m, 2 H), 4.08 (m, 2H), 4.61 (m, 1 H), 7.09 (m, 2 H), 7.35 (m, 5 H), 7.49 (t, J=9.1 Hz, 1H), 7.59 (t, J=9.0 Hz, 1 H), and 7.81 (3 H).

(G) Homophenylalanyl-Nα-Methylornithinol 2-Naphthyl EtherTrifluoroacetate

Treatment of Boc-homophenylalanyl-Nγ-Boc-Nα-methylornithinol 2-naphthylether (F) (60 mg) with trifluoroacetic acid (Procedure E) affordedproduct as a white solid (63 mg); HPLC (method A): ¹ H NMR (400 MHz, D₂O) δ 1.77 (m, 4 H), 2.01 (m, 1 H), 2.15 (m, 1 H), 2.75 (m, 2 H), 2.89(s, 3 H), 3.11 (m, 2 H), 4.26 (dd, J=10.8; 3.2 Hz, 1 H), 4.34 (t, J=10.8Hz, 1 H), 4.55 (m, 1 H), 5.08 (m, 1 H), 7.09 (m, 2 H), 7.35 (m, 5 H),7.49 (t, J=7.6 Hz, 1 H), 7.59 (t, J=7.2 Hz, 1 H), 7.75 (d, J=9.2 Hz, 1H), 7.82 (d, J=8.4 Hz, 1 H), and 7.89 (d, J=8.4 Hz, 1 H).

Example 103 O-Benzylseryl-Nα-Methylornithinol 2-Naphthyl EtherTrifluoroacetate

(A) Boc-O-Benzylseryl-Nγ-Boc-Nα-Methylornithinol 2-Naphthyl Ether

Using Procedure D, crude Nγ-Boc-Nα-methylornithinol 2-naphthyl ether (37mg) and Boc-O-benzylserine (62 mg) was coupled to afford product (62 mg)as a colorless oil: ¹ H NMR (400 MHz, CDCl₃) δ 1.49 (broad s, 21 H),1.77 (m, 1 H), 3.08 (s, 3 H), 3.17 (s, 2 H), 3.64 (m, 2 H), 4.05 (dd,J=13.0; 3.8 Hz, 1 H), 4.14 (dd, J=13.0; 7.3 Hz, 1 H),4.52 (m, 3 H), 4.91(m, 1 H), 7.05 (m, 2 H), 7.20 (m, 3 H), 7.32 (m, 3 H), 7.41 (m, 1 H),and 7.69 (m, 3 H); mass spectrum (ES+) m/e 636 (M+1).

(B) O-Benzylseryl-Nα-Methylornithinol 2-Naphthyl Ether Trifluoroacetate

Boc-O-benzylseryl-Nγ-Boc-Nα-methylornithinol 2-naphthyl ether, aftertreatment with trifluoroacetic acid, afforded titled compound as a whitesolid: HPLC (method C); ¹ H NMR (400 MHz, CDCl₃) δ 1.52 (m, 1 H), 1.63(m, 2 H), 1.78 (m, 1 H), 2.82 (s, 3 H), 2.88 (m, 2 H), 3.86 (broad s, 4H), 4.47 (m, 2 H), 4.61 (m, 1 H), 5.05 (m, 1 H), 6.86 (d, J=10.8 Hz,III), 6.92 (s, 1H1), 7.10 (m, 3 H), 7.18 (m, 2 H), 7.30 (t, J=8.0 Hz, 1H), 7.39 (t, J=8.1 Hz, 1 H), 7.61 (t, J=8.7 Hz, 2 H), and 7.75 (d, J=9.0Hz, 1 H); mass spectrum (ES+) m/e 436 (M+1).

Example 104 Tyrosyl-Nα-Methylornithinol 2-Naphthyl EtherTrifluoroacetate

(A) Boc-Tyrosyl-Nγ-Boc-Nα-Methylornithinol 2-Naphthyl Ether

Using Procedure D, crude Nγ-Boc-Nα-methylornithinol 2-naphthyl ether(133 mg) and Boc-tyrosine (222 mg) is coupled to afford intermediate(125 mg) as a glassy solid: ¹ H NMR (400 MHz, CDCl₃) δ 1.44 (m, 21 H),1.63 (m, 1 H), 2.75 (s, 3 H), 3.15 (m, 2 H), 3.92 (m, 2 H), 4.84 (m, 1H), 5.06 (m, 1 H), 6.62 (d, J=10.2 Hz, 2 H), 7.08 (m, 4 H), 7.35 (m, 1H), 7.42 (m, 1 H), 7.66 (d, J=10.9 Hz, 1 H), and 7.75 (m, 3 H).

(B) Tyrosyl-Nα-Methylornithinol 2-Naphthyl Ether Trifluoroacetate

Boc-Tyrosyl-Nγ-Boc-Nα-methylorrithinol 2-naphthyl ether (125 mg) wastreated with trifluoroacetic acid (Procedure E) to afford titled product(100 mg) as a white solid: ¹ H NMR (400 MHz, D₂ O) δ 1.77 (m, 4 H), 2.78(m, 3 H), 3.08 (m, 4 H), 4.06 (dd, J=11.4; 3.6 Hz, 1 H), 4.17 (dd,J=11.5; 8.4 Hz, 1 H), 4.72 (t, J=7.8 Hz, 1 H), 4.89 (m, 1 H), 6.65 (d,J=10.8 Hz, 2 H), 7.17 (d, J=10.8 Hz, 2 H), 7.22 (dd, J=9.6; 1.1 Hz, 1H), 7.34 (s, 1 H), 7.51 (t, J=8.4 Hz, 1 H), 7.61 (t, J=8.4 Hz, 1 H),7.94 (m, 3 H).

Example 105 Phenylalanyl-Nα-Methylornithinol (4-Methoxy-2-naphthyl)etherTrifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-phenylalanine and Nγ-Boc-Nα-methylornithinol(4-methoxy-2-naphthyl)ether.

Example 106 Tyrosyl-Nα-Methylornithinol (4-Methoxy-2-naphthyl)etherTrifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-tyrosine and Nγ-Boc-Nα-methylornithinol(4-methoxy-2-naphthyl)ether.

Example 107 Phenylalanyl-Nα-Benzylornithinol (4-Methoxy-2-naphthyl)etherTrifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-phenylalanine and Nγ-Boc-Nα-benzylornithinol(4-methoxy-2-naphthyl)ether.

Example 108 Tyrosyl-Nα-Ethylornithinol (4-Methoxy-2-naphthyl)etherTrifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-tyrosine and Nγ-Boc-Nα-ethylomnithinol(4-methoxy-2-naphthyl)ether.

Example 109 4-Fluorohomophenylalanyl-Nα-Methylornithinol2-Naphthyl-Ether Trifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-4-fluorohomophenylalanine andNγ-Boc-Nα-methylomithinol 2-naphthyl ether.

Example 110 4-Fluorohomophenylalanyl-Nα-Methylornithinol 2-QuinolinylEther Trifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-4-fluorohomophenylalanine andNγ-Boc-Nα-methylornithinol 2-quinolinyl ether.

Example 111 Homophenylalanyl-Nα-Methylornithinol 3-Quinolinyl EtherTrifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-homophenylalanine and Nγ-Boc-Nα-methylornithinol3-quinolinyl ether.

Example 112 3-Fluorotyrosyl-Nα-Methylornithinol 4-Quinolinyl EtherTrifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-3-fluorotyrosine and Nγ-Boc-Nα-methylornithinol4-quinolinyl ether.

Example 113 Homophenylalanyl-Nα-(4-Methoxybenzyl)ornithinol 3-QuinolinylEther Trifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-homophenylalanine andNγ-Boc-Nα-(4-methoxybenzyl)ornithinol 3-quinolinyl ether.

Example 114 Trytophane-Nα-Methylornithinol 3-Quinolinyl EtherTrifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-trytophane and Nγ-Boc-Nα-methylornithinol3-quinolinyl ether.

Example 115 2,4-Dichlorophenylalanyl-Nα-Methylornithinol(3,4-Dimethylphenyl)ether Trifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-2,4-dichlorophenylalanine andNγ-Boc-Nα-methylornithinol (3,4-dimethylphenyl)ether.

Example 116 D-(2-Naphthyl)alanyl-Nα-Methylornithinol(3,4-Dimethoxyphenyl)ether Trifluoroacetate

This was prepared, as described in Example 104, except the startingmaterials were Boc-β-(2-naphthyl)alanine and Nγ-Boc-Nα-methylornithinol(3,4-dimethylphenyl)ether.

Example 117 Homophenylalanyl-Nα-Methylargininol 2-Naphthyl EtherTrifluoroacetate

(A) Nω,Nω'-Bis-Boc-Nα-Methylargininol 2-Naphthyl Ether

This compound is prepared in three steps fromNα-benzyl-Nγ-Boc-Nα-methylargininol. First, the Boc protecting group isremoved using trifluoroacetic acid, then the amine salt isguanidiny-lated with N,N'-bis-Boc-1-guanylpyrrazole. Removal of thebenzyl group was accomplished by catalytic hydrogenation with 5%palladium-on-carbon: ¹ H NMR (400 MHz, CDCl₃) δ 1.48 (s, 18 H), 1.73 (m,4 H), 2.48 (s, 3 H), 2.95 (m, 1 H), 3.47 (m, 2 H), 4.04 (dd, J=10.9; 6.0Hz, 1 H), 4.13 (dd, J=11.0; 2.7 Hz, 1 H), 7.15 (m, 2 H), 7.35 (t, J=8.4Hz, 1 H), 7.44 (t, J=8.4 Hz, 1 H), and 7.94 (m, 3 H).

(B) Boc-Homophenylalanyl-Nω,Nω∝-BisBoc-Nα-Methylargininol 2-NaphthylEther

Using Procedure D, Nω,Nω'-Bis-Boc-Nα-methylargininol 2-naphthyl etherwas coupled with Boc-homophenylalanine to afford a white solid: ¹ H NMR(400 MHz, D₂ O) δ 1.47 (m, 29 H), 1.66 (m, 1 H), 1.76 (m, 1 H), 1.95 (m,1 H), 2.04 (m, 1 H), 2.73 (m, 2 H), 2.85 (s, 3 H), 4.11 (m, 2 H), 4.62(m, 1 H), 5.14 (m, 1 H), 6.99-7.16 (m, 7 H), 7.49 (t, ¹ H), 7.59 (t, 1H), and 7.64-7.77 (m, 3 H).

(C) Homophenylalanyl-Nα-Methylargininol 2-Naphthyl EtherTrifluoroacetate

This compound is obtained by treatment ofBoc-homophenylalanyl-Nω,Nω'-Bis-Boc-Nα-methylargininol 2-naphthyl etherwith trifluoroacetic acid, followed by HPLC purification.

Example 118 N-(C-Amidino)homophenylalanyl-Nα-Methylargininol 2-NaphthylEther Trifluoroacetate

Homophenylalanyl-N.sub.α -methylornithinol 2-naphthyl ether (41 mg) andN,N'-bis-Boc-1-guanylpyrrazole (19 mg) was coupled to affordN-(bis-Boc-C-amidino)homophenyl-alanine-NωNω'-bis-Boc-Nα-methylargininol2-naphthyl ether (56 mg). Deprotection of the intermediate by ProcedureE, followed by HPLC purification afforded titled product as a whitesolid: ¹ H NMR (400 MHz, D₂ O) δ 1.65 (m, 2 H), 1.72 (m, 2 H), 1.93 (m,1 H), 2.13 (m, 1 H), 2.78 (m, 2 H), 2.92 (2 s, 3 H, rotamers), 3.29 (m,2 H), 4.27 (m, 1 H), 4.38 (t, J=13.0 Hz, 1 H), 4.53 (m, 1 H), 5.03 (m, 1H), 7.12 (m, 2 H), 7.37 (m, 4 H), 7.52 (m, 1 H), 7.60 (m, 1 H), and7.81-7.96 (m, 3 H).

Example 119 Homophenylalanyl-Nα-Methylornithinethiol 2-BenzothiazolylThioether Trifluoroacetate

(A) O-(tert-Butyldimethylsilyl)-Nα-Benzyl-Nγ-Boc-Nα-Methylornithinol

A solution of Nα-benzyl-Nγ-Boc-Nα-methylornithinol (560 mg),dimethylformamide (1.5 ml), and t-butyldimethylsilyl chloride (330 mg),triethylamine (290 μl) and 4-(N,N-dimethylamino)-pyridine (21 mg) wasstirred at 0° C., under nitrogen atmosphere, for 1 hr. Then the mixturewas stirred, at 25° C., for 10 hrs and then poured into water (20 ml)and extracted with dichloro-methane (2×20 ml). The combined organicphase was washed with water and brine, and dried over sodium sulfate.Evaporation of the solvent afforded titled compound (690 mg) as gas aclear solid: ¹ H NMR (400 MHz, CDCl₃) δ 0.06 (s, 6 H), 0.91 (s, 9 H),1.43 (s, 9 H), 1.45-1.73 (m, 4 H), 2.22 (s, 3 H), 2.68 (m, 1 H), 3.12(m, 2 H), 3.62 (dd, J=10.4; 5.2 Hz, 1 H), 3.68 (d, J=13.6 Hz, 1 H), 3.78(m, 2 H), and 7.30 (m, 5 H).

(B) O-(tert-Butyldimethylsilyl)-Nγ-Boc-Nα-Methylornithinol

Reduction ofO-(tert-butyldimethylsilyl)-Nα-benzyl-Nγ-Boc-Nα-methylornithinol (A)(690 mg) afforded titled compound (485mg) which was used in thesubsequent reaction: ¹ H NMR (400 MHz, CDCl₃) δ 0.09 (s, 6 H), 0.92 (s,9 H), 1.39-1.68 (m, 13 H), 2.40 (s, 3 H), 2.48 (m, 1 H), 3.12 (m, 2 H),3.46 (dd, J=10.0; 6.4 Hz, 1 H), and 3.62 (dd, J=9.6; 4.0 Hz, 1 H).

(C)Boc-Homophenylalanyl-O-(tert-butyldimethylsilyl)-Nγ-Boc-Nα-Methylornithinol

Using Procedure D, coupling ofO-(tert-butyldimethylsilyl)-Nγ-Boc-Nα-methylornithinol (B) (485 mg) andBoc-homophenylalanine (590 mg) afforded titled compound (767 mg) as aglassy solid: ¹ H NMR (400 MHz, CDCl₃) δ 0.01 (s, 6 H), 0.84 (s, 9 H),1.39-1.55 (broad s, 22 H), 1.84 (m, 1 H), 1.97 (m, 1 H), 2.69 (m, 2 H),2.78 (s, 3 H), 3.09 (m, 2 H), 3.58 (d, J=5.7 Hz, 2 H), 4.58 (m, 2 H),7.20 (m, 3 H), and 7.26 (m, 2 H).

(D) Boc-Homophenylalanyl-Nγ-Boc-Nα-Methylornithinol

A mixture of tetrabutylammonium fluoride (2.8 ml of 1M sol. intetrahydrofuran),Boc-homophenylalanyl-O-(tert-butyldimethylsilyl)-Nγ-Boc-Nα-methylornithinol(C) (576 mg), and dry tetrahydrofuran (5 ml) was stirred at 0° C. for 1hr. The reaction mixture is poured into ethyl acetate and worked up,including purification by flash chromatography (5%methanol/dichloromethane) to yield desired product (402 mg) as a thickoil: ¹ H NMR (400 MHz, CDCl₃) δ 1.44 (broad s, 22 H), 1.86 (m, 1 H),2.00 (m, 1 H), 2.79 (m, 4 H) 3.09 (m, 2 H), 3.43-3.69 (m, 2 H), 4.45 (m,1 H), 4.63 (m, 1 H), and 7.19-7.32 (m, 5 H).

(E) Homophenylalanyl-Nα-Methylornithinethiol 2-Benzothiazolyl ThioetherTrifluoracetate

This was prepared in a two step sequence. A solution ofBoc-homophenylalanine-Nγ-Boc-Nα-methylornithinol (D) (392 mg) and2-mercaptobenzothiazole (267 mg) in dry tetrahydrofiran (9 ml) wascooled to 0° C. under nitrogen atmosphere. Then, a solution oftriphenylphosphine (1.04 g), anhydrous tetrahydrofuran (1 ml), anddiethyl azodicarboxylate (620 μl ) was added. The reaction mixture wasstirred for 1.5 h at 0° C., concentrated in vacuo, and purified by flashchromatography (30% ethyl acetate/hexane) to affordBoc-homophenylalanyl-Nγ-Boc-Nα-methylornithinethiol 2-benzothiazolylthio-ether (270 mg). This intermediate was deprotected by Procedure E toafford titled product (75 mg): HPLC [20 to 40% gradient(acetonitrile/0.1% TFA) over 60 min, retention time=42.96 min]; ¹ H NMR(400 MHz, D₂ O) δ 1.49 (m, 2 H), 1.83 (m, 2 H), 1.98 (m, 1 H), 2.18 (m,1 H), 2.79 (m, 2 H), 2.82 (s, 3 H), 3.09 (m, 2H), 3.57 (dd, J=14.8; 11.2Hz, 1 H), 3.70 (dd, J=14.8; 4.0 Hz, 1 H), 4.46 (m, 1 H), 5.01 (m, 1 H),7.17 (m, 2 H), 7.38 (m, 3 H), 7.49 (t, J=8.0 Hz, 1 H), 7.61 (t, J=7.2Hz, 1 H), 7.91 (d, J=8.0 Hz, 1 H), and 7.94 (d, J=8.4 Hz, 1 H).

Example 120 Phenylalanyl-Nα-Methylornithinethiol 3-Quinolinyl ThioetherTrifluoroacetate

This was prepared, as described in Example 119, except the startingmaterials were Boc-phenylalanine and Nα-methylomithinethiol 3-quinolinylthioether.

Example 121 Homophenylalanyl-Nα-Ethylornithinethiol 3-QuinolinylThioether Trifluoroacetate

This was prepared, as described in Example 119, except the startingmaterials were Boc-homophenylalanine and Nα-ethylornithinethiol3-quinolinyl thioether.

Example 122 Phenylalanyl-Nα-Methylornithinethiol 2-Quinolinyl ThioetherTrifluoroacetate

This was prepared, as described in Example 119, except the startingmaterials were Boc-phenylalanine and Nα-methylornithinethiol2-quinolinyl thioether.

Example 123 Trytophan-Nα-Methylornithinethiol 4-Quinolinyl ThioetherTrifluoroacetate Example 1244-Chlorophenylalanyl-Nα-Methylornithinethiol 2-Quinolinyl ThioetherTrifluoroacetate

This was prepared, as described in Example 119, except the startingmaterials were Boc-trytophane and Nα-methylornithinethiol 4-quinolinylthioether.

Example 124 4-Chlorophenylalanyl-Nα-Methylornithinethiol 2-QuinolinylThioether Trifluoroacetate

This was prepared, as described in Example 119, except the startingmaterials were Boc-4-chlorophenylalanine and Nα-methylornithinethiol2-quinolinyl thioether.

Example 125 Homophenylalanyl-Nα-Methylornithinethiol 2-BenzimidazolylThioether Trifluoroacetate

This was prepared, as described in Example 119, except the startingmaterials were Boc-homophenylalanine and Nα-methylornithinethiol2-benzimidazolyl thioether.

Example 126 Homophenylalanyl-Nα-Methyllysinethiol 2-BenzimidazolylThioether Trifluoroacetate

This was prepared, as described in Example 119, except the startingmaterials were Boc-homophenylalanine and Nα-methyllysinethiol2-benzimidazolyl thioether.

Example 127 Tyrosyl-Nα-Methyllysinethiol 2-Benzimidazolyl ThioetherTrifluoroacetate

This was prepared, as described in Example 119, except the startingmaterials were Boc-tyrosine and Nα-methyllysinethiol 2-benzimidazolylthioether.

The embodiments described herein are not meant to be limiting to theinvention. Those skilled in the art will appreciate that the inventioncan be practiced using various microbial strains and with various modesof contacting a microbe with an efflux pump inhibitor or or treating aninfection and that the compounds of this invention can be prepared by avariety of synthetic methods, all within the scope of the invention.

Other embodiments are within the following claims.

What we claim is:
 1. A method for treating a microbial infection in ananimal, comprising administering to an animal suffering from saidinfection an antimicrobial agent and an efflux pump inhibitor in anamount sufficient to reduce efflux pump activity,wherein said effluxpump inhibitor increases the susceptibility of said microbe to saidantimicrobial agent, and wherein said efflux pump inhibitor has thechemical structure of structure 1 below: ##STR7## wherein M* is(CH₂)_(n) (n=0, 1, or 2) P* is CH₂, carbonyl (C═O), or thiocarbonyl(C═S) S* is CH₂, CH(OH), NH, O, or SO_(t) (t=0, 1, or 2); R is H, loweralkyl, branched alkyl, fluoroalkyl, perfluoroalkyl, carboxyalkyl,hydroxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n)NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n)N═CNR^(b) R^(c), wherein n=1, 2, 3, or 4, and R^(a), R^(b), and R^(c)are independely H, lower alkyl, phenyl, substituted phenyl, benzyl,cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or--CH═CH--; R¹ is H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, carboxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, arylalkyl, arylalkyl, thienylalkyl, furylalkyl,pyridylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3, or 4 and R^(a),R^(b), and R^(c) are indpendently H, lower alkyl, phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) (or R^(b) +R^(c)) is (CH₂)₂₋₃ or--CH═CH--; R² is H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)-pyridyl, benzofuranyl, benzothienyl, indolyl, benzimidazolyl,benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl, furylalkyl,pyridylalkyl, benzofuranylalkyl, benzothienylalkyl, indolylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n)N═CNR^(b) R^(c), wherein n=1, 2, 3, or 4 and R^(a), R^(b), and R^(c) areindependently H, lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro,or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--, except that ifM* is (CH₂)_(n) (n═0) and P* is carbonyl (C═O) and S* is NH then R² isdifferent from H; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,quinoxalinylalkyl, quinazolinylalkyl, benzimidazolylalkyl,benzothiazolylalkyl, or benzoxazolylalkyl.
 2. The method of claim 1,wherein said efflux pump inhibitor has Structure 2, ##STR8## wherein Ris H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, hydroxyalkyl, aryl, monosubstituted aryl, disubstitutedaryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b)R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c),wherein n=1, 2, 3, or 4 and R^(a), R^(b), and R^(c) are independently H,lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b)or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--;R¹ is H, lower alkyl, branchedalkyl, fluoroalkyl, perfluoroalkyl, carboxyalkyl, hydroxyalkyl, aryl,2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c),(CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c),(CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c), whereinn=1, 2, 3, or 4 and R^(a), R^(b) or R^(c) are independently H, loweralkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) orR^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R² is [H,] lower alkyl, branchedalkyl, fluoroalkyl, perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl, benzothienyl, indolyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, benzofuranylalkyl, benzothienylalkyl,indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3, or 4 and R^(a),R^(b) and R^(c) are independently H, lower alkyl, phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or--CH═CH--; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,isoquinolinyl, quinoxalinylalkyl, quinazolinylalkyl,benzimidazolylalkyl, benzothiazolylalkyl, or benzoxazolylalkyl.
 3. Themethod of claim 1, wherein said efflux pump inhibitor has Structure 3,##STR9## wherein R is H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, carboxy-alkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl,2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3,or 4 and R^(a), R^(b) and R^(c) are independently H, lower alkyl,phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(b)+R^(c) is (CH₂)₂ -₃ or --CH═CH--;R¹ is H, lower alkyl, branched alkyl,fluoroalkyl, perfluoroalkyl, carboxy-alkyl, hydroxyalkyl, aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c),(CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c),(CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), whereinn=1, 2, 3 or 4 and R^(a), R^(b) and R^(c) are independently H, loweralkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) orR^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R² is H, lower alkyl, branchedalkyl, fluoroalkyl, perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl, benzothienyl, indolyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, benzofuranylalkyl, benzothienylalkyl,indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3, or 4 and R^(a),R^(b) and R^(c) are independently H, lower alkyl, phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or--CH═CH--; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, arnidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,isoquinolinyl, quinoxalinylalkyl, quinazolinylalkyl,benzirnidazolylalkyl, benzothiazolylalkyl, or benzoxazolylalkyl.
 4. Themethod of claim 1, wherein said efflux pump inhibitor has structure 4,##STR10## wherein S* is CH₂, CH(OH), NH, O, or SO_(t) (t=0,1, or 2);R isH, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR^(b) R^(c), wherein n=1, 2, 3, or 4 and R^(a), R^(b) and R^(c) areindependently H, alkyl, phenyl, benzyl, cyano, hydroxy, or nitro, orR^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R¹ is H, alkyl,branched alkyl, fluoroalkyl, perfluoroalkyl, carboxyalkyl, aryl,2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c),(CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c),(CH₂)_(n) C═(NR^(a))NR_(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c), whereinn=1, 2, 3, or 4 and R^(a), R^(b) and R^(c) are independently H, loweralkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) orR^(b) ═R^(c) is (CH₂)₂₋₃ or --CH═CH--; R² is H, lower alkyl, branchedalkyl, fluoroalkyl, perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl, benzothienyl, indolyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, benzofuranylalkyl, benzothienylalkyl,indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), (CH₂)_(n) N═NR^(b) R^(c), wherein n=1, 2, 3, or 4 and R^(a),R^(b) and R^(c) are independently H, lower alkyl, phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or--CH═CH--; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,quinoxalinylalkyl, quinazolinylalkyl, benzimidazolylalkyl,benzothiazolylalkyl, or benzoxazolylalkyl.
 5. A method for prophylactictreatment of an animal, comprising administering to an animal at risk ofa microbial infection an antimicrobial agent and an efflux pumpinhibitor, wherein said efflux pump inhibitor increases thesusceptibility of a microbe to said antimicrobial agent, andwherein saidefflux pump inhibitor has the chemical structure of structure 1 below:##STR11## wherein M* is (CH₂)_(n) (n=0, 1, or 2) P* is CH₂, carbonyl(C═O), or thiocarbonyl (C═S) S* is CH₂, CH(OH), NH, O, or SO_(t) (t=0, 1or 2); R is H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n)N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4, and R^(a), R^(b), and R^(c) areindependently H, lower alkyl, phenyl, substituted phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or--CH═CH--; R¹ is H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, carboxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, arylalkyl, arylalkyl, thienylalkyl, furylalkyl,pyridylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a),R^(b), and R^(c) are indpendently H, lower alkyl, phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) (or R^(b) +R^(c)) is (CH₂)₂₋₃ or--CH═CH--; R² is H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or ³ -)furyl, or 2-(3- or4-)-pyridyl, benzofuranyl, benzothienyl, indolyl, benzirnidazolyl,benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl, furylalkyl,pyridylalkyl, benzofuranylalkyl, benzothienylalkyl, indolylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n)N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a), R^(b), and R^(c) areindependently H, lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro,or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--, except that ifM* is (CH₂)_(n) (n=0) and P* is carbonyl (C═O) and S* is NH then R² isdifferent from H; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,quinoxalinylalkyl, quinazolinylalkyl, benzimidazolylalkyl,benzothiazolylalkyl, or benzoxazolylalkyl.
 6. The method of claim 5,wherein said efflux pump inhibitor has Structure 2, ##STR12## wherein Ris H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, hydroxyalkyl, aryl, monosubstituted aryl, disubstitutedaryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b)R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c),wherein n=1, 2, 3 or 4 and R^(a), R^(b), and R^(c) are independently H,lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b)or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--;R¹ is H, lower alkyl, branchedalkyl, fluoroalkyl, perfluoroalkyl, carboxyalkyl, hydroxyalkyl, aryl,2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b)R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c),wherein n=1, 2, 3 or 4 and R^(a), R^(b) or R^(c) are independently H,lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b)or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R² is [H,] lower alkyl,branched alkyl, fluoroalkyl, perfluoroalkyl, aryl, 2-(or 3-)thienyl,2-(or 3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl, benzothienyl,indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, benzofuranylalkyl,benzothienylalkyl, indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═R^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) Re, wherein n=1, 2, 3 or 4 andR^(a), R^(b) and R^(c) are independently H, lower alkyl, phenyl, benzyl,cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or--CH═CH--; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,isoquinolinyl, quinoxalinylalkyl, quinazolinylalkyl,benzimidazolylalkyl, benzothiazolylalkyl, or benzoxazolylalkyl.
 7. Themethod of claim 5, wherein said efflux pump inhibitor has Structure 3,##STR13## wherein R is H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, carboxy-alkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl,2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3or 4 and R^(a), R^(b) and R^(c) are independently H, lower alkyl,phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(b)+R^(c) is (CH₂)₂₋₃ or --CH═CH--;R¹ is H, lower alkyl, branched alkyl,fluoroalkyl, perfluoroalkyl, carboxy-alkyl, hydroxyalkyl, aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)NR^(b) R^(c), (CH₂)_(n)NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR_(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3or 4 and R^(a), R^(b) and R^(c) are independently H, lower alkyl,phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(b)+R^(c) is (CH₂)₂₋₃ or --CH═CH--; R² is H, lower alkyl, branched aikyl,fluoroalkyl, perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)-pyridyl, benzofuiranyl, benzothienyl, indolyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, benzofuranylalkyl, benzothienylalkyl,indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(R^(a))NR^(b) R^(c),(CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c),(CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a), R^(b) andR^(c) are independently H, lower alkyl, phenyl, benzyl, cyano, hydroxy,or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; W is(alpha-aminoacyl)amido, arninoalkyl, amnino, azaheterocycles,substituted azaheterocycles, hydroxy, alkoxy, alkylthio, guanidino,amidino, or halogen; and X is aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, tetrahydronaphthyl, indanyl, quinolinyl,isoquinolinyl, quinoxalinyl, quinazolinyl, benzimidazolyl,benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl, furylalkyl,pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl, isoquinolinyl,quinoxalinylalkyl, quinazolinylalkyl, benzimidazolylalkyl,benzothiazolylalkyl, or benzoxazolylalkyl.
 8. The method of claim 5,wherein said efflux pump inhibitor has structure 4, ##STR14## wherein S*is CH₂, CH(OH), NH, O, or SO_(t) (t=0, 1, or 2);R is H, lower alkyl,branched alkyl, fluoroalkyl, perfluoroalkyl, carboxyalkyl, hydroxyalkyl,aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b)R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c),wherein n=1, 2, 3 or 4 and R^(a), R^(b) and R^(c) are independently H,alkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) orR^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R¹ is H, alkyl, branched alkyl,fluoroalkyl, perfluoroalkyl, carboxyalkyl, aryl,2-(or 3-)thienyl, 2-(or3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl,pyridylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a),R^(b) and R^(c) are independently H, lower alkyl, phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or--CH═CH--; R² is H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)-pyridyl, benzofuranyl, benzothienyl, indolyl, benzimidazolyl,benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl, furylalkyl,pyridylalkyl, benzofuranylalkyl, benzothienylalkyl, indolylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR_(b) R_(c), wherein n=1, 2, 3 or 4 and R^(a), R^(b) and R^(c) areindependently H, lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro,or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; W is(alpha-aminoacyl)amido, aminoalkyl, amino, azaheterocycles, substitutedazaheterocycles, hydroxy, alkoxy, alkylthio, guanidino, amidino, orhalogen; and X is aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)pyridyl, tetrahydronaphthyl, indanyl, quinolinyl, isoquinolinyl,quinoxalinyl, quinazolinyl, benzimidazolyl, benzothiazolyl,benzoxazolyl, arylalkyl, thienylalkyl, fuirylalkyl, pyridylalkyl,quinolinylalkyl, isoquinolinylalkyl, quinoxalinylalkyl,quinazolinylalkyl, benzimidazolylalkyl, benzothiazolylalkyl, orbenzoxazolylalkyl.
 9. The method of any of claims 1 or 5, wherein saidanimal is a mamnmal.
 10. A method of enhancing the antimicrobialactivity of an antimicrobial agent against a microbe, comprisingcontacting said microbe with said antimicrobial agent and an efflux pumpinhibitor in an amount effective to inhibit an efflux pump in saidmicrobe,wherein said efflux pump inhibitor has the chemical structure ofstructure 1 below: ##STR15## wherein M* is (CH₂)_(n) (n=0, 1, or 2) P*is CH₂, carbonyl (C═O), or thiocarbonyl (C═S) S* is CH₂, CH(OH), NH, O,or SO_(t) (t=0, 1, or 2); R is H, lower alkyl, branched alkyl,fluoroalkyl, perfluoroalkyl, carboxyalkyl, hydroxyalkyl, aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c),(CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c),(CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), whereinn=1, 2, 3 or 4, and R^(a), R^(b), and R^(c) are independently H, loweralkyl, phenyl, substituted phenyl, benzyl, cyano, hydroxy, or nitro, orR^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R¹ is H, loweralkyl, branched alkyl, fluoroalkyl, perfluoroalkyl, carboxyalkyl, aryl,2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b)R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c),wherein n=1, 2, 3 or 4 and R^(a), R^(b), and R^(c) are indpendently H,lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b)(or R^(b) +R^(c)) is (CH₂)₂₋₃ or --CH═CH--; R² is H, lower alkyl,branched alkyl, fluoroalkyl, perfluoroalkyl, aryl, 2-(or 3-)thienyl,2-(or 3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl, benzothienyl,indolyl, benzirnidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, benzofuranylalkyl,benzothienylalkyl, indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3or 4 and R^(a), R^(b), and R^(c) are independently H, lower alkyl,phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(b)+R^(c) is (CH₂)₂₋₃ or --CH═CH--, except that if M* is (CH₂)_(n) (n=0)and P* is carbonyl (C═O) and S* is NH, then R² is different from H; W is(alpha-aminoacyl)amido, aminoalkyl, amino, azaheterocycles, substitutedazaheterocycles, hydroxy, alkoxy, alkylthio, guanidino, amidino, orhalogen; and X is aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)pyridyl, tetrahydronaphthyl, indanyl, quinolinyl, isoquinolinyl,quinoxalinyl, quinazolinyl, benzimidazolyl, benzothiazolyl,benzoxazolyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,quinolinylalkyl, isoquinolinylalkyl, quinoxalinylalkyl,quinazolinylalkyl, benzimidazolylalkyl, benzothiazolylalkyl, orbenzoxazolylalkyl.
 11. The method of claim 10, wherein said efflux pumpinhibitor has Structure 2, ##STR16## wherein R is [H,] lower alkyl,branched alkyl, fluoroalkyl, perfluoroalkyl, carboxyalkyl, hydroxyalkyl,aryl, monosubstituted aryl, disubstituted aryl, 2-(or 3-)thienyl, 2-(or3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl,pyridylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a),R^(b), and R^(c) are independently H, lower alkyl, phenyl, benzyl,cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(a) +R^(c) is (CH₂)₂₋₃ or--CH═CH--;R¹ is H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, carboxyalkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl,2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl,thienylalkyl,furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or4 and R^(a), R^(b) or R^(c) are independently H, lower alkyl, phenyl,benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is(CH₂)₂₋₃ or --CH═CH--; R² is H, lower alkyl, branched alkyl,fluoroalkyl, perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)-pyridyl, benzofuranyl, benzothienyl, indolyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, benzofuranylalkyl, benzothienylalkyl,indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a),R^(b) and R^(c) are independently H, lower alkyl, phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or--CH═CH--; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,isoquinolinyl, quinoxalinylalkyl, quinazolinylalkyl,benzimidazolylalkyl, benzothiazolylalkyl, or benzoxazolylalkyl.
 12. Themethod of claim 10, wherein said efflux pump inhibitor has Structure 3,##STR17## wherein R is H, lower alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, carboxy-alkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl,2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(NR_(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3or 4 and R^(a), R^(b) and R^(c) are independently H, lower alkyl,phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) ═R^(b) or R^(b)═R^(c) is (CH₂)₂₋₃ or --CH═CH--;R¹ is H, lower alkyl, branched alkyl,fluoroalkyl, perfluoroalkyl, carboxy-alkyl, hydroxyalkyl, aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c),(CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(R^(a))NR^(b) R^(c),(CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c), whereinn=1, 2, 3 or 4 and R^(a), R^(b) and R^(c) are independently H, loweralkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) orR^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R² is H, lower alkyl, branchedalkyl, fluoroalkyl, perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl, benzothienyl, indolyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, benzofuranylalkyl, benzothienylalkyl,indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a),R^(b) and R^(c) are independently H, lower alkyl, phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or--CH═CH--; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,isoquinolinyl, quinoxalinylalkyl, quinazolinylalkyl,benzimidazolylalkyl, benzothiazolylalkyl, or benzoxazolylalkyl.
 13. Themethod of claim 10, wherein said efflux pump inhibitor has structure 4,##STR18## wherein S* is CH₂, CH(OH), NH, O, or SO_(t) (t=0, 1 or 2);R isH, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, hydroxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a), R^(b) and R^(c) areindependently H, alkyl, phenyl, benzyl, cyano, hydroxy, or nitro, orR^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R¹ is H, alkyl,branched alkyl, fluoroalkyl, perfluoroalkyl, carboxyalkyl, aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c),(CH₂)_(n) NHC═R^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c),(CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c), whereinn=1, 2, 3 or 4 and R^(a), R^(b) and R^(c) are independently H, loweralkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) orR^(b) ═R^(c) is (CH₂)₂₋₃ or --CH═CH--; R² is H, lower alkyl, branchedalkyl, fluoroalkyl, perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl, benzothienyl, indolyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, benzofuranylalkyl, benzothienylalkyl,indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a),R^(b) and R^(c) are independently H, lower alkyl, phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) or R^(b) R^(c) is (CH₂)₂₋₃ or--CH═CH--; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,quinoxalinylalkyl, quinazolinylalkyl, benzimidazolylalkyl,benzothiazolylalkyl, or benzoxazolylalkyl.
 14. The method of any ofclaims 1, 5, or 10, wherein said microbe is a bacterium.
 15. The methodof claim 14, wherein said bacterial infection involves a bacteriumselected from the group consisting of Pseudomonas aeruginosa,Pseudomonas fluorescens, Pseudomonas acidovorans, Pseudomonasalcaligenes, Pseudomonas putida, Stenotrophomonas maltophilia,Burkholderia cepacia, Aeromonas hydrophilia, Escherichia coli,Citrobacter freundii, Salmonella typhimurium, Salmonella typhi,Salmonella paratyphi, Salmonella enteritidis, Shigella dysenteriae,Shigella flexneri, Shigella sonnei, Enterobacter cloacae, Enterobacteraerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Serratiamarcescens, Francisella tularensis, Morganella morganii, Proteusmirabilis, Proteus vulgaris, Providencia alcalifaciens, Providenciarettgeri, Providencia stuartii, Acinetobacter calcoaceticus,Acinetobacter haemolyticus, Yersinia enterocolitica, Yersinia pestis,Yersinia pseudotuberculosis, Yersinia intermedia, Bordetella pertussis,Bordetella parapertussis, Bordetella bronchiseptica, Haemophilusinfluenzae, Haemophilus parainfluenzae, Haemophilus haemolyticus,Haemophilus parahaemolyticus, Haemophilus ducreyi, Pasteurellamultocida, Pasteurella haemolytica, Branhamella catarrhalis,Helicobacter pylori, Campylobacter fetus, Campylobacter jejuni,Campylobacter coli, Borrelia burgdorferi, Vibrio cholerae, Vibrioparahaemolyticus, Legionella pneumophila, Listeria monocytogenes,Neisseria gonorrhoeae, Neisseria meningitidis, Gardnerella vaginalis,Bacteroides fragilis, Bacteroides distasonis, Bacteroides 3452A homologygroup, Bacteroides vulgatus, Bacteroides ovalus, Bacteroidesthetaiotaomicron, Bacteroides uniformis, Bacteroides eggerthii,Bacteroides splanchnicus, Clostridium difficile, Mycobacteriumtuberculosis, Mycobacterium avium, Mycobacterium intracellulare,Mycobacterium leprae, Corynebacterium diphtheriae, Corynebacteriumulcerans, Streptococcus pneumoniae, Streptococcus agalactiae,Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium,Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcussaprophyticus, Staphylococcus intermedius, Staphylococcus hyicus subsp.hyicus, Staphylococcus haemolyticus, Staphylococcus hominis,Staphylococcus saccharolyticus.
 16. The method of any of claims 1, 5, or10, wherein said microbial infection is a bacterial infection and saidantimicrobial agent is an antibacterial agent.
 17. The method of claim16, wherein said antibacterial agent is a quinolone.
 18. The method ofclaim 16, wherein said antibacterial agent is a tetracycline.
 19. Themethod of claim 16, wherein said antibacterial agent is a β-lactam. 20.The method of claim 16, wherein said antibacterial agent is acoumermycin.
 21. The method of claim 16, wherein said antibacterialagent is chloramphenicol.
 22. The method of claim 16, wherein saidantibacterial agent is a glycopeptide.
 23. The method of claim 16,wherein said antibacterial agent is an aminoglycoside.
 24. The method ofclaim 16, wherein said antibacterial agent is a macrolide.
 25. Themethod of claim 16, wherein said antibacterial agent is a rifamycin. 26.A pharmaceutical composition effective for treatment of an infection ofan animal by a microbe, comprising an efflux pump inhibitor and apharmaceutically acceptable carrier,wherein said efflux pump inhibitorhas the chemical structure of structure 1 below: ##STR19## wherein M* is(CH₂)_(n) (n=0, 1 or 2) P* is CH₂, carbonyl (C═O), or thiocarbonyl (C═S)S* is CH₂, CH(OH), NH, O, or SO_(t) (t=0, 1 or 2); R is H, lower alkyl,branched alkyl, fluoroalkyl, perfluoroalkyl, carboxyalkyl, hydroxyalkyl,aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b)R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c),wherein n=1, 2, 3 or 4, and R^(a), R^(b), and R^(c) are independently H,lower alkyl, phenyl, substituted phenyl, benzyl, cyano, hydroxy, ornitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R¹ isH, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)pyridyl, arylalkyl, arylalkyl, thienylalkyl, furylalkyl,pyridylalkyl, (CH₂)NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c),(CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or(CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a), R^(b), andR^(c) are indpendently H, lower alkyl, phenyl, benzyl, cyano, hydroxy,or nitro, or R^(a) +R^(b) (or R^(b) +R^(c)) is (CH₂)₂₋₃ or --CH═CH--; R²is H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl, aryl,2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)-pyridyl,benzofliranyl, benzothienyl, indolyl, benzimnidazolyl, benzothiazolyl,benzoxazolyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,benzofuranylalkyl, benzothienylalkyl, indolylalkyl, (CH₂)_(n) NR^(b)R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c),wherein n=1, 2, 3 or 4 and R^(a), R^(b), and R^(c) are independently H,lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b)or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--, except that if M* is (CH₂)_(n)(n=O) and P* is carbonyl (C═O) and S* is NH then R² is different from H;W is (alpha-aminoacyl)amido, aminoalkyl, amino, azaheterocycles,substituted azaheterocycles, hydroxy, alkoxy, alkylthio, guanidino,amidino, or halogen; and X is aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or2-(3- or 4-)pyridyl, tetrahydronaphthyl, indanyl, quinolinyl,isoquinolinyl, quinoxalinyl, quinazolinyl, benzimidazolyl,benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl, furylalkyl,pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl, quinoxalinylalkyl,quinazolinylalkyl, benzimidazolylalkyl, benzothiazolylalkyl, orbenzoxazolylalkyl.
 27. The pharmaceutical composition of claim 26,wherein said efflux pump inhibitor has Structure 2, ##STR20## wherein Ris H, lower alkyl, branched alkyl, fluoroalkyl, perfluoroalkyl,carboxyalkyl, hydroxyalkyl, aryl, monosubstituted aryl, disubstitutedaryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b)R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n) N═CNR^(b) R^(c),wherein n=1, 2, 3 or 4 and R^(a), R^(b), and R^(c) are independently H,lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b)or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--;R¹ is H, lower alkyl, branchedalkyl, fluoroalkyl, perfluoroalkyl, carboxyalkyl, hydroxyalkyl, aryl,2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl,arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b)R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b)R^(c), (CH₂)nC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c), whereinn=1, 2, 3 or 4 and R^(a), R^(b) or R^(c) are independently H, loweralkyl, phenyl, benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) orR^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R² is [H,] lower alkyl, branchedalkyl, fluoroalkyl, perfluoroalkyl, aryl, 2-(or 3-)thienyl, 2-(or3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl, benzothienyl, indolyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, benzofuranylalkyl, benzothienylalkyl,indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b)R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b)R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a),R^(b) and R^(c) are independently H, lower alkyl, phenyl, benzyl, cyano,hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or--CH═CH--; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,isoquinolinyl, quinoxalinylalkyl, quinazolinylalkyl,benzimidazolylalkyl, benzothiazolylalkyl, or benzoxazolylalkyl.
 28. Thepharmaceutical composition of claim 26, wherein said efflux pumpinhibitor has Structure 3, ##STR21## wherein R is H, lower alkyl,branched alkyl, fluoroalkyl, perfluoroalkyl, carboxy-alkyl,hydroxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n)NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n)N═CNR^(R) ^(c), wherein n=1, 2, 3 or 4 and R^(a), R^(b) and R^(c) areindependently H, lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro,or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--;R¹ is H, loweralkyl, branched alkyl, fluoroalkyl, perfluoroalkyl, carboxy-alkyl,hydroxyalkyl, aryl, 2-(or 3-)thienyl, 2-(or 3-)furyl, or 2-(3- or4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n)NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), or (CH₂)_(n)N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a), R^(b) and R^(c) areindependently H, lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro,or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R² is H, loweralkyl, branched alkyl, fluoroalkyl, perfluoroalkyl, aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl,benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, benzofuranylalkyl,benzothienylalkyl, indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or4 and R^(a), R^(b) and R^(c) are independently H, lower alkyl, phenyl,benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is(CH₂)₂₋₃ or --CH═CH--; W is (alpha-aminoacyl)arnido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,isoquinolinyl, quinoxalinylalkyl, quinazolinylalkyl,benzimidazolylalkyl, benzothiazolylalkyl, or benzoxazolylalkyl.
 29. Thepharmaceutical composition of claim 1, wherein said efflux pumpinhibitor has. structure 4, ##STR22## wherein S* is CH₂, CH(OH), NH, O,or SO_(t) (t=0, 1, or 2);R is H, lower alkyl, branched alkyl,fluoroalkyl, perfluoroalkyl, carboxyalkyl, hydroxyalkyl, aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, arylalkyl,thienylalkyl, furylalkyl, pyridylalkyl, (CH₂)_(n) NR^(b) R^(c),(CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c),(CH₂)_(n) C═(R^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1,2, 3 or 4 and R^(a), R^(b) and R^(c) are independently H, alkyl, phenyl,benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is(CH₂)₂₋₃ or --CH═CH--; R¹ is H, alkyl, branched alkyl, fluoroalkyl,perfluoroalkyl, carboxyalkyl, aryl,2-(or 3-)thienyl, 2-(or ³ -)furyl, or2-(3- or 4-)pyridyl, arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl,(CH₂)_(n) NR^(b) R^(c), (CH₂)_(n) NHC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n) C═(NR^(a))NR^(b) R^(c), (CH₂)_(n)N═CNR^(b) R^(c), wherein n=1, 2, 3 or 4 and R^(a), R^(b) and R^(c) areindependently H, lower alkyl, phenyl, benzyl, cyano, hydroxy, or nitro,or R^(a) +R^(b) or R^(b) +R^(c) is (CH₂)₂₋₃ or --CH═CH--; R² is H, loweralkyl, branched alkyl, fluoroalkyl, perfluoroalkyl, aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)-pyridyl, benzofuranyl,benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl,arylalkyl, thienylalkyl, furylalkyl, pyridylalkyl, benzofuranylalkyl,benzothienylalkyl, indolylalkyl, (CH₂)_(n) NR^(b) R^(c), (CH₂)_(n)NHC═(R^(a))NR^(b) R^(c), (CH₂)_(n) SC═(NR^(a))NR^(b) R^(c), (CH₂)_(n)C═(NR^(a))NR^(b) R^(c), (CH₂)_(n) N═CNR^(b) R^(c), wherein n=1, 2, 3 or4 and R^(a), R^(b) and R^(c) are independently H, lower alkyl, phenyl,benzyl, cyano, hydroxy, or nitro, or R^(a) +R^(b) or R^(b) +R^(c) is(CH₂)₂₋₃ or --CH═CH--; W is (alpha-aminoacyl)amido, aminoalkyl, amino,azaheterocycles, substituted azaheterocycles, hydroxy, alkoxy,alkylthio, guanidino, amidino, or halogen; and X is aryl, 2-(or3-)thienyl, 2-(or 3-)furyl, or 2-(3- or 4-)pyridyl, tetrahydronaphthyl,indanyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, arylalkyl, thienylalkyl,furylalkyl, pyridylalkyl, quinolinylalkyl, isoquinolinylalkyl,quinoxalinylalkyl, quinazolinylalkyl, benzimidazolylalkyl,benzothiazolylalkyl, or benzoxazolylalkyl.
 30. The pharmaceuticalcomposition of claim 26, wherein said microbe is a bacterium.
 31. Thepharmaceutical composition of claim 26, further comprising anantimicrobial agent.
 32. The pharmaceutical composition of claim 31,wherein said microbe is a bacterium.
 33. The pharmaceutical compositionof claim 32, wherein said antimicrobial agent is an antibacterial agent.